Methods and related compositions for reduction of fat

ABSTRACT

Compositions and methods useful in the reduction of localized fat deposits in patients in need thereof using pharmacologically active detergents are disclosed. The pharmacologically active detergent compositions can additionally include anti-inflammatory agents, analgesics, dispersion or anti-dispersion agents and pharmaceutically acceptable excipients. The pharmacologically active detergent compositions are useful for treating localized accumulations of fat including, for example, lower eyelid fat herniation, lipodystrophy and fat deposits associated with cellulite and do not require surgical procedures such as liposuction.

CROSS-REFERENCE

The present application claims priority to U.S. Provisional ApplicationSer. No. 60/572,879 filed May 19, 2004, which is incorporated herein byreference in its entirety.

FIELD OF THE INVENTION

The present invention is related to compositions and methods useful forthe non-surgical removal of localized fat accumulation. Specifically,the present invention is related to pharmacologically active detergentcompositions than are suitable for injection directly into a treatmentsite of a patient in need of fat removal without the need for surgicalintervention.

BACKGROUND OF THE INVENTION

Numbers appearing in parentheses at the end of a sentence refer tospecific references cited at the conclusion of this specificationimmediately before the claims.

Formulations containing phosphatidylcholine and bile salts(phosphatidylcholine bile salt formulations, PBF) are increasingly beingutilized to treat localized fat accumulation (1-8). Several open labelclinical studies have reported promising results using injections ofPBFs for the treatment of localized fat accumulation, including lowereyelid fat herniation and “buffalo hump” lipodystrophy (1-3).

Phosphatidylcholine is a natural phospholipid that is an essentialcomponent of cell membranes and is important for normal cellularmembrane composition and repair. Phosphatidylcholine is also the majordelivery form of the essential nutrient choline. Choline itself is aprecursor in the synthesis of the neurotransmitter acetylcholine, themethyl donor betaine and phospholipids, including phosphatidylcholineand sphingomyelin among others. Phosphatidylcholine is also involved inthe hepatic export of very-low-density lipoproteins.

Bile salts have been used to improve the aqueous solubility ofphosphatidylcholine and more recently, medications like amphotericin B,Taxol®, and diazepam (9-14). Highly purified phosphatidylcholine can becombined with the secondary bile salt sodium deoxycholate, ananti-microbial, benzyl alcohol, and water to form a stable, mixedmicelle preparation that can be rapidly sterilized and used forintravenous administration (12). Pharmaceutical preparations of thismixture, known as Essentiale® and Lipostabil®, are marketed in othercountries for treatment of liver disease and hyperlipidemia,respectively (12,15).

Rittes first reported that injections of a PBF into subcutaneous fatreduced infraorbital fat herniation (1). Since then, physicians havebeen using the pharmaceutical preparations or similar, compounded PBFs,to treat lower eyelid fat herniation, as well as fat deposits on thethighs, abdomen, upper back, chin, and arms (2,3,5). These PBFs oftenlack the dl-alpha-tocopherol (vitamin E), B-vitamins, and adenosinemonophosphate variably found in Essentiale® and Lipostabil® (2,16).

Phosphatidylcholine formulations are associated with localized burningsensations, erythema, transient urticaria and variable degrees ofpruritus all of which usually resolve within a few days. More serioussequelae of ulceration and pain have also been seen. An infectiousgranulomatous reaction has been reported in the thigh of a patient atthe site of multiple phosphatidylcholine injections (7). Increaseddosages of injected phosphatidylcholine have paralleled side effectsseen with large doses of oral and intravenous formulations ofLipostabil® and include nausea, diarrhea, abdominal pain and syncope.

The mechanism whereby phosphatidylcholine-containing formulation causereduction of subcutaneous fat deposits is unknown but several mechanismshave been proposed (4). The first is that phosphatidylcholine couldreduce the size of lipocytes by stimulating lipase activity.Alternatively, the PBFs have been postulated to function as a detergentthat emulsifies lipocyte cell membranes. Detergents have been used inmedicine for decades, specifically, as sclerosing agents commonly usedin sclerotherapy (American College of Phlebology, 2003). Detergentspossess unique polar and non-polar chemical properties which facilitatesemulsification of insoluble substances by reducing surface tension attheir interface (17). In fact, laboratory detergents like Triton® X-100and Empigen® BB are commonly used to disrupt the lipid bilayer of cellmembranes (10,18-21). Two major components of the PBFs,phosphatidylcholine and sodium deoxycholate, have these unique chemicalproperties and therefore have been used independently as detergents oremulsifying agents (9,18,20-25).

Surgical and non-surgical procedures for improving appearance haveincreased in prevalence as populations age and gain weight. Liposuctionis one of the most popular cosmetic surgery procedures and involves thesurgical removal of fat deposits using suction and optionally assistedby solutions to assist in fat removal. Liposuction, also known aslipoplasty or suction lipectomy, is a surgical procedure that removesfat through an incision in the skin through which a cannula is inserted.The cannula is connected to a suction source and the unwanted fat isaspirated through the cannula and discarded. Liposuction is performedunder general or local anesthesia, depending on the amount and locationof the fat to be removed.

The most commonly used forms of liposuction additionally use fluidinjection methodologies wherein a medicated solution containing amixture of salts, an anesthetic and a vasoconstrictor, is infused intothe treatment site prior to aspiration of the fat tissue. The medicatedsolution helps the fat be removed more easily, reduces blood loss andprovides anesthesia both during and after surgery.

In an example of adjuvant solutions for liposuction, a United Statespatent filed on Apr. 22, 1997 and issued as U.S. Pat. No. 5,891,083 onApr. 6, 1999 by Capella and Capella teaches liposuction and a carriersolution containing a compound for an improved surgical procedure forremoving subcutaneous fat. In one embodiment the Capella patentdiscloses the compound is an enzyme, particularly lipase or colipase.The enzyme is added to a carrier such as saline solution to provide alipolysis solution. In another embodiment of the invention, Capellateaches emulsifying agents such as bile salts may also be beneficial incombination or as the primary active compound added to the solution. Inevery embodiment of the Capella invention, the lipolysis solution isadministered for a period of time before liposuction to allow for thesolution to infiltrate the fat tissue. Nowhere in Capella is the use ofa lipolysis solution alone disclosed as a non-surgical means forremoving fat from the body. In all examples and specific embodimentsdisclosed in Capella, liposuction is used as a surgical procedure forfat removal and lipase and bile salts are provided as an adjuvant toliposuction.

However, liposuction and other surgical methods of fat removal areassociated with significant adverse events including temporary bruising,swelling, numbness, soreness and burning sensation, risk of infection,pigmentation changes; the formation of fat clots or blood clots whichcan migrate to the lungs and cause death, excessive fluid loss, whichcan lead to shock or fluid accumulation that must be drained, frictionburns or other damage to the skin or nerves or perforation injury to thevital organs. Additionally, liposuction requires a recovery time of oneto two weeks wherein the patient cannot work or perform certain dailyactivities. Moreover, because surgical procedures such as liposuctionrequire local and occasionally general anesthesia, significantanesthesia-related risks are associated with surgical fat removal.

Therefore it would be desirable to have a method of removing localizedfat accumulations that does not require surgery or prolonged recoverytime and has fewer adverse side effects than currently availablemethods.

SUMMARY OF THE INVENTION

The present invention provides methods and kits for reducingsubcutaneous fat deposits. In one aspect, the invention contemplateskits having a first container comprising a pharmacologically activedetergent and less than 5% w/v phosphatidylcholine, as well as writteninstructions for reducing subcutaneous fat deposits in a mammal withoutthe use of surgery. Preferably, the kits herein may be used to reducefat deposits in a variety of mammals such as, for example, a human, ahorse, a dog, or a cat. In some embodiments the mammal is a human.

In some preferred embodiments, the first container has a total volume ofless than 500 ml and/or is provided as an injectable formulation. Inother preferred embodiments, the first container may contain a % w/v ofdetergent greater than the % w/v of phosphatidylcholine or may containno phosphatidylcholine. In one preferred embodiment, the presentinvention provides the detergent at a concentration above its criticalmicellar concentration (CMC). The kits may comprise a variety ofpharmacologically active detergents such as, for example, a lipophilicdetergent, a hydrophilic detergent, an ionic detergent, a non-ionicdetergent, a glyceride, a bile salt, and a zwitterionic detergent. In amore preferred embodiment, the active detergent is a bile salt, mostpreferably sodium deoxycholate. A first container in the kit herein may,in some embodiments include less than 3 g detergent. In otherembodiments, a first container in the kit herein may include more than0.0002 g detergent. In any of the embodiments herein the first containermay further include a second detergent.

Preferably, the first container may further comprise a secondtherapeutic agent such as, for example, an anti-microbial agent, avasoconstrictor, an anti-thrombotic agent, an anti-coagulation agent, asuds-depressant, an anti-inflammatory agent, an analgesic, a dispersionagent, an anti-dispersion agent, a penetration enhancer, a steroid, atranquilizer, a muscle relaxant, and an anti-diarrhea agent. In someembodiments the second therapeutic agent is an analgesic, anti-microbialagent, or an anti-inflammatory agent. More preferably, the secondtherapeutic agent is an analgesic, or most preferably lidocain. Inanother embodiment, the kit provides a second container comprising thesecond therapeutic agent as described herein.

One embodiment of the present invention contemplates a kit herein forreducing fat deposits under the eye, chin, or arm, as well as thebuttock, calf, back, thigh, ankle, or stomach of a mammal. In anotherembodiment, the kit may reduce specific types of fat deposits such as,for example, eyelid fat herniation, lipomas, lipodystrophy, buffalo humplipodystrophy, or fat deposits associated with cellulite.

In a second aspect, the present invention provides methods for reducingsubcutaneous fat deposits in a mammal without surgery by administering aunit dose comprising an effective amount of a pharmacologically activedetergent and less than 5% w/v phosphatidylcholine. In one embodiment,the methods do not include the step of actively removing the detergent.The methods may be used to reduce fat deposits in a variety of mammalsas described herein.

In another embodiment, the method includes the step of administeringless than 500 ml of a solution comprising an effective amount ofdetergent and less than 5% w/v phosphatidylcholine. In some preferredembodiments, the unit dose is administered locally and/or is repeated atleast twice. In other preferred embodiments, the unit dose has a greater% w/v of detergent than % w/v of phosphatidylcholine and/or has aconcentration of detergent above its CMC. The methods may use a varietyof pharmacologically active detergents as described herein. In a morepreferred embodiment, the active detergent in the unit dose is a bilesalt, most preferably sodium deoxycholate.

Preferably, the methods herein may further comprise the administrationof a second detergent and/or a second therapeutic agent. The methods mayinclude the administration of a variety of second therapeutic agents asdescribed herein. More preferably, the second therapeutic agent is ananalgesic, most preferably lidocain.

One other embodiment contemplates one or more methods described hereinto reduce fat deposits under the eye, chin, or arm, as well as thebuttock, calf, back, thigh, ankle, or stomach of a mammal. In anotherembodiment, the methods may reduce specific types of fat deposits suchas, for example, eyelid fat herniation, lipomas, lipodystrophy, buffalohump lipodystrophy, or fat deposits associated with cellulite.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the molecular structure of (a) phosphatidylcholine (b)sodium deoxycholate and (c) benzyl alcohol.

FIG. 2 depicts the effects of phosphatidylcholine bile formulation (PCFormula, PBF) and sodium deoxycholate alone on cultured cell viabilityaccording to the teachings of the present invention: (a) MTS assaymeasuring viability of keratinocytes exposed to the PC Formula andsodium deoxycholate alone; (b) Lactate dehydrogenase (LDH) assaymeasuring LDH release by cells exposed to the PC Formula and sodiumdeoxycholate alone.

FIG. 3 depicts the effects of PBF and sodium deoxycholate alone onprimary porcine fat tissue according to the teachings of the presentinvention: (a) MTS assay producing purple pigment, indicating livingcells, in fat specimens treated with the PBS buffer as negative control(−Cont), sodium deoxycholate alone (DC), the PBF (PC), and Triton®detergent as positive control (+Cont); (b) A comparison of fat cellviability between the different treatments.

FIG. 4 depicts calcein fluorescence in fat specimens treated with sodiumdeoxycholate alone (DC), PBF (PC), Triton® detergent as positive control(+Cont), and PBS buffer as negative control (−Cont) according to theteachings of the present invention.

FIG. 5 depicts light microscopy of porcine skin biopsies after treatmentwith compositions made according to the teachings of the presentinvention revealing (a) control lipocytes and (b) lipocytes after PBFinjection (H&E, original magnification, ×20); (c) control lipocytes and(d) lipocytes after injection of sodium deoxycholate alone (H&E,original magnification, ×10); (e) control muscle and (f) muscle afterinjection of phosphatidylcholine alone (H&E, original magnification,×10); (g) fat after injection with Empigen® detergent (H&E, originalmagnification, ×20).

FIG. 6 depicts a lipoma removed from a patient two days after injectionwith deoxycholate according to the teachings of the present invention:(a) gross pathology and (b) histology (H&E, original magnification,×20).

FIG. 7 depicts a kit for reducing a subcutaneous fat accumulation.

DETAILED DESCRIPTION OF THE INVENTION

The present invention addresses the problem of localized fataccumulation in mammals by providing a non-surgical method for reducingfat deposits by administration of fat-solubilizing concentrations of oneor more detergents in pharmaceutically acceptable formulations.

Based on phosphatidylcholine's role as an emulsifier in bile and its usein the treatment of hyperlipidemia, phosphatidylcholine has beenpostulated as the active ingredient in PBFs (1, 2, 21, 25, 27). Thedetergents such as bile salts in these prior art compositions were addedmerely to disperse or solubilize the presumed active ingredient, PC.However, to date, there are no published reports supporting this theory.The present inventors have unexpectedly demonstrated that the bile saltwas actually the active agent for localized fat emulsification.

Numbers appearing in parentheses at the end of a sentence refer tospecific references cited at the conclusion of this specificationimmediately before the claims. All of the references cited herein arehereby incorporated by reference in their entirety for all purposes.

Phosphatidylcholine is a natural phospholipid that is an essentialcomponent of cell membranes and is important for normal cellularmembrane composition and repair. Phosphatidylcholine is also the majordelivery form of the essential nutrient choline. Choline itself is aprecursor in the synthesis of the neurotransmitter acetylcholine, themethyl donor betaine and phospholipids, including phosphatidylcholineand sphingomyelin among others. Phosphatidylcholine is also involved inthe hepatic export of very-low-density lipoproteins.

Bile salts have been used to improve the aqueous solubility ofphosphatidylcholine and more recently, medications like amphotericin B,Taxol™, and diazepam (9-14). Highly purified phosphatidylcholine can becombined with the secondary bile salt sodium deoxycholate, ananti-microbial, benzyl alcohol, and water to form a stable, mixedmicelle preparation that can be rapidly sterilized and used forintravenous administration (12). Pharmaceutical preparations of thismixture, known as Essentiale® and Lipostabil®, are marketed in othercountries for treatment of liver disease and hyperlipidemia,respectively (12, 15).

Physicians have been using pharmaceutical preparations or compoundedPBFs to treat lower eyelid fat herniation, as well as fat deposits onthe thighs, abdomen, upper back, chin, and arms (2, 3, 5). These PBFsoften lack the dl-alpha-tocopherol (vitamin E), B-vitamins, andadenosine monophosphate variably found in Essentiale® and Lipostabil®(2, 16).

Liposuction is one of the most popular cosmetic surgery procedures andinvolves the surgical removal of fat deposits using suction andoptionally assisted by solutions to assist in fat removal. Liposuction,also known as lipoplasty or suction lipectomy, is a surgical procedurethat reduces fat through an incision in the skin through which a cannulais inserted. The cannula is connected to a suction source and theunwanted fat is aspirated through the cannula and discarded. Liposuctionis performed under general or local anesthesia, depending on the amountand location of the fat to be reduced. Such infusions consisted of largevolumes of solution and are often forced out of the patient prior to orduring a liposuction procedure. See U.S. Pat. No. 5,891,083.

The use of liposuction and/or other surgical methods of fat removal areassociated with significant adverse events including temporary bruising,swelling, numbness, soreness and burning sensation, risk of infection,pigmentation changes; the formation of fat clots or blood clots whichcan migrate to the lungs and cause death, excessive fluid loss, whichcan lead to shock or fluid accumulation that must be drained, frictionburns or other damage to the skin or nerves or perforation injury to thevital organs. Additionally, liposuction requires a recovery time of oneto two weeks wherein the patient cannot work or perform certain dailyactivities. Moreover, because surgical procedures such as liposuctionrequire local and occasionally general anesthesia, significantanesthesia-related risks are associated with surgical fat removal.

While meeting with some success, prior techniques and compositions havemet with certain limitations. Therefore it would be desirable to have amethod of reducing localized fat accumulations that does not requiresurgery or prolonged recovery time and has fewer adverse side effectsthan currently available methods.

The present invention relates to the use of one or morepharmacologically active detergents (e.g., bile salts) to reducesubcutaneous fat accumulations in a mammal by administering suchformulation locally to a target site.

Among detergents, bile salts are particularly potent solubilizers oflipid bilayer membranes (9, 20, 21, 23, 28). All biologic cell membranesare composed of the same bilipid structure, and are therefore subject tosolubilization by detergents (10, 19, 34). Solubilization of cellmembranes by a detergent involves distribution of the detergent betweenlipid bilayers, destabilization of the bilayer, disintegration, andsubsequent formation of mixed micelles (composed of detergent and cellmembrane lipid) (10, 19,21). Bile salts, and other detergents, decreasesurface tension at the border of immiscible materials and allows thebreakdown of large aggregates into smaller and smaller particles. Intissue, these agents dissolve cell membranes and cause cell lysis. Aninflammatory response is generated, causing the body to remove thedetergent solubilized material.

For this reason, the present inventors compared sodium deoxycholate withthe complete PBF using a simple, quantitative assay measuring cellviability (FIG. 2 a). It is not possible to isolate and test purephosphatidylcholine because it is insoluble in aqueous solutions unlessit is combined with substances like bile salts (12). Phosphatidylcholineis highly soluble in ethanol, methanol, chloroform, and other organicsolvents, yet these agents can damage lipid bilayers (29-31). Inpreliminary experiments, there was no difference in cell lysis andhistology between pure, isolated PC and the ethanol used to dissolve it.Although benzyl alcohol, one of the components of the PC formula, hasbeen shown to affect the fluidity of cell membranes, it is a not adetergent, and therefore, its limited quantity in the formula hasnegligible lytic effects on cell membranes (32, 33).

Because penetration into intact tissues may be likely a limiting factor,cell cultures were used to determine the dilutions of the reagents (PBFand deoxycholate) necessary to affect cells. Deoxycholate profoundlydecreased the viability of cultured cells approximately equal to thecomplete PBF (FIG. 2 a). This finding was reproduced in tissue byexposing porcine fat to PBF and deoxycholate (FIG. 3). These resultssupport the unexpected observation that sodium deoxycholate plays amajor, active role in the PBF.

The present invention is based on the use of detergent action ofdisrupting cell membrane to reduce subcutaneous fat deposits. Membranelysis in cultured cells was measured using a lactate dehydrogenase (LDH)assay and within tissue using calcein, a fluorescent marker retained incells with intact cell membranes. The LDH assay measures the activity ofLDH, which is a cytosolic enzyme released when cells are lysed. Both thePBF- and deoxycholate-treated cell cultures demonstrated aconcentration-dependent increase in cell lysis (FIG. 2 b). Moreover, thedirect lytic effects observed in cultured cells treated with theseagents suggest activity independent of endogenous lipase. Calcein waslost in the fat specimens exposed to the PBF, deoxycholate, and Triton®X-100, a known laboratory detergent (FIG. 4). This finding confirmedthat disruption of cell membranes occurs in fresh tissue exposed to boththe PBF and deoxycholate.

Comparing the effects of the PBF to deoxycholate in cell culture led tothe surprising result that deoxycholate caused similar loss of cellviability, but less cell lysis. These differences may be concentrationdependent or there may be synergistic effects betweenphosphatidylcholine and deoxycholate within the formula. Nonetheless,the data demonstrate that, at concentrations similar to those usedclinically, deoxycholate and the PBF had similar effects on tissuehistology and cell viability. Taken together, these data unexpectedlydemonstrate that deoxycholate acts as the active component in the priorart PBF.

In order to illustrate the effect of detergents on tissue histology,fresh porcine skin was injected with PBF, deoxycholate, andwell-characterized laboratory detergents (FIG. 5). All reagents causedsignificant disruption of lipocyte organization compared to PBSinjection (control). These results were similarly observed within muscleand connective tissue. Rapid dissolution of cell borders by the testsubstances and the similarity of their effects to well characterizeddetergents substantiate that the PBF and deoxycholate function asdetergents. The limitation with this experimental model is that it doesnot reveal the true sequelae that occur after injection into livingtissue. It is apparent from clinical reports that a brisk inflammatoryresponse, evident as erythema and edema, occurs after injection (1-3).Repeated inflammation can potentially lead to fibrosis, especially aftermultiple injections. Fibrosis has been reported in several patients whodeveloped firm nodules at injection sites after PBF administration thateventually resolve over several months (35).

Histologic findings reveal that the injectable PBF and deoxycholatealone cause architectural disruption in fat and muscle, but had noapparent affect on the epidermis, dermis, or adnexae (FIG. 5). However,Empigen® BB, a potent laboratory detergent, had profound histologiceffects on dermal collagen (connective tissue). Alternatively, fat andmuscle can be more sensitive to detergent treatment than these otherstructures at the tested concentrations (similar to those used inclinical practice).

Through a series of laboratory experiments utilizing fresh tissuespecimens and cell cultures, the present inventors have demonstratedthat the prior art PBF popularly used in subcutaneous injections for fatdissolution works primarily by causing non-specific lysis of cellmembranes. Cell membranes are constituents of all tissue types;specifically, the present inventor demonstrated that these detergentscause solubilization of fat, muscle and connective tissue. Therefore thepresent inventors concluded that sodium deoxycholate, the bile saltcomponent of the formula used to dissolve the phosphatidylcholine, wasthe major active ingredient of these prior art formulations. Thisconclusion is supported by the fact that pharmacologically activedetergents, such as bile salts are potent solubilizers of cellmembranes. Moreover, the mechanism of the PBF and sodium deoxycholate infat dissolution is likely detergent action.

Compositions

In an embodiment of the present invention, a medical composition ofbiologically compatible detergents includes one or morepharmacologically active detergents and pharmaceutically acceptableexcipients in an aqueous vehicle. In particular, it is within the scopeof the present invention that pharmacologically active detergentsincluding bile salts are used to dissolve fat.

In one embodiment, the present invention relates compositions comprise,consist essentially of, or consist of one or more pharmacologicallyactive detergents in an effective amount to reduce subcutaneous fat.

Pharmacologically active detergents that can be used in embodiments ofthe present invention include, but are not limited to, lipophilicdetergents (whether ionic or non-ionic), hydrophilic detergents (whetherionic or non-ionic), ionic detergents, non-ionic detergents,zwitterionic detergents, glycerides, and bile salts.

Non-limiting examples of lipophilic detergents include, inter alia,alcohols; polyoxyethylene alkylethers; fatty acids, bile acids; glycerolfatty acid esters; acetylated glycerol fatty acid esters; lower alcoholfatty acids esters; polyethylene glycol fatty acid esters; polyethyleneglycol glycerol fatty acid esters; polypropylene glycol fatty acidesters; polyoxyethylene glycerides; lactic acid derivatives ofmono/diglycerides; propylene glycol diglycerides; sorbitan fatty acidesters; polyoxyethylene sorbitan fatty acid esters;polyoxyethylene-polyoxypropylene block copolymers; transesterifiedvegetable oils; sterols; sterol derivatives; sugar esters; sugar ethers;sucroglycerides; polyoxyethylene vegetable oils; polyoxyethylenehydrogenated vegetable oils; reaction mixtures of polyols and at leastone member of the group consisting of fatty acids, glycerides, vegetableoils, hydrogenated vegetable oils, and sterols; and mixtures thereof.

Non-limiting examples of non-ionic lipophilic detergents include, interalia, alkylglucosides; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; polyoxyethylene alkyl ethers; polyoxyethylenealkylphenols; polyethylene glycol fatty acids esters, polyethyleneglycol glycerol fatty acid esters; polyoxyethylene sorbitan fatty acidesters; polyoxyethylene-polyoxypropylene block copolymers; polyglycerolfatty acid esters; polyoxyethylene glycerides; polyoxyethylene sterols,derivatives, and analogues thereof; polyoxyethylene vegetable oils;polyoxyethylene hydrogenated vegetable oils; reaction mixtures ofpolyols and at least one member of the group consisting of fatty acids,glycerides, vegetable oils, hydrogenated vegetable oils, and sterols;tocopherol polyethylene glycol succinates; sugar esters; sugar ethers;sucroglycerides; and mixtures thereof.

Non-limiting examples of ionic hydrophilic detergents include, interalia, alkyl ammonium salts, bile acids and salts, analogues, andderivatives thereof; fatty acid derivatives of amino acids, carnitines,oligopeptides, and polypeptides; glyceride derivatives of amino acids,oligopeptides, and polypeptides; acyl lactylates; mono-, diacetylatedtartaric acid esters of mono-, diglycerides; succinoylatedmonoglycerides; citric acid esters of mono-, diglycerides; alginatesalts; propylene glycol alginate; lecithins and hydrogenated lecithins;lysolecithin and hydrogenated lysolecithins; lysophospholipids andderivatives thereof, phospholipids and derivatives thereof; salts ofalkylsulphates; salts of fatty acids; sodium docusate; and mixturesthereof.

Non-limiting examples of ionic detergents include, but not limited to,cholate, sodium deoxycholate, sodium dodecylsulfate and C-16 TAB. Inpreferred embodiment, a non-limiting example of an ionic detergentuseful in an embodiment of the present invention is sodium deoxycholate.

Non-limiting examples of non-ionic detergents include, but not limitedto, Brij 35, n-alkyl PEO monoether such as, polyoxylethylen(20)cetylether, Lubrol PX, Lubrol WX, nonidet P-40, n-alkyl phenyl PEO such as,octylphenolpoly(ethyleneglycolether)n10, andoctylphenolpoly(ethyleneglycolether)n7, tetramethylbutylphenyl PEO,n-octylglucoside, octyl-thioglucopyranoside, tween-80 and tween-20, andalkylaryl polyether alcohol (Triton® X-100).

Non-limiting examples of zwitterionic detergents include, but notlimited to, 3-[(3-cholamidopropyl)dimthylammonio]propane-sulfonate(CHAPS), N-tetradecyl-N,N-dimethyl-3-ammoniu-1-propanesulfonate, cholicacid sulfobetaine, lauryldimethylbetaine (Empigen® BB) and zwittergent3-14.

Non-limiting examples of glycerides include, inter alia, mono-, di- ortri-glycerides. Such triglycerides include, inter alia, vegetable oils,fish oils, animal fats, hydrogenated vegetable oils, partiallyhydrogenated vegetable oils, synthetic triglycerides, modifiedtriglycerides, fractionated triglycerides, and mixtures thereof.

Non-limiting examples of bile salts include steroids having 1-3 hydroxylgroups and a five carbon atom side chain terminating in a carboxylgroup, which can be conjugated to glycine or taurine.

Additional examples of bile salts include salts of cholate, deoxycholic,cholic, chenodeoxycholic, 7-alpha-dehydroxylate, chenodeoxycholic,lithocholic, ursodeoxycholic, dihydroxy- and trihydroxy- and taurine orglycine conjugates of any of the above. Preferably a bile salt of theinvention is sodium deoxycholate.

Table 1 below illustrates several detergents contemplated by the presentinvention, their monomeric molecular weight of these detergents asmonomers, and their critical micellar concentration (CMCs), which is theminimum concentration at which the detergent is predominantly in theform of micelles.

TABLE 1 Micellar Molecular Molecular CMC in Weight Weight H2O DetergentName (AMU) (AMU) (M) Anionic Cholate 430 4300  1.4 × 10−2 Deoxycholate415-432 4200   5 × 10−3 Sodium dodecyl sulfate 288 18000  8.3 × 10−3cationic C16-TAB 365 62000   1 × 10−3 Amphoteric (Zwiterionic) Cholicacid-sulfobetaine 615 6150   4 × 10−3 Cholic acid-sulfobetaine 631 6940  8 × 10−3 Lysophophatidylcholine 495 92000   7 × 10−6 Zwitergent 3-14364 30000   3 × 10−4 Non-Ionic Brij 35 1225 49000   9 × 10−5polyoxylethylen(20)cetyl ether 1120 82000  7.7 × 10−5 Lubrol PX 58264000   1 × 10−4 Nonidet P-40 603 90000   3 × 10−4 Octylphenolpoly 64790000  0.2 × 10−3 (ethyleneglycolether)n10 Octylphenolpoly 515  0.2 ×10−3 (ethyleneglycolether)n7 n-Octylglucoside 292 8000 14.5 × 10−3Octyl-thioglucopyranoside 308   9 × 10−3 Tween-80 1310 76000  1.2 × 10−5Tween-20 1228  6.0 × 10−5

Preferably, the concentration of the one or more pharmacologicallyactive detergents in a composition herein is such that it is atapproximately the CMC concentration (i.e. +/−5 mM), or at aconcentration that is above the CMC level, such as more than 1%, 5%, 10%15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 99%, 150%, 200%, 400%, 800%, 1600%, 3200%, 6800%, 13,600%, 27,200%,or 54,400%, above the CMC concentration level.

In some embodiments, a concentration of the one or more of thepharmacologically active detergents in a composition is less than 20%,19%, 18%, 17%, 16%, 15%,14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%,0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%,0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%,0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v or v/v.

In some embodiments, a concentration of the one or more of thepharmacologically active detergents in a composition is greater than20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%,17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25%15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%, 12.75%,12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25%10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25%7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%,4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%,0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%,0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%,0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v, or v/v.

In some embodiments, a concentration of the one or more of thepharmacologically active detergents in a composition is in the rangefrom approximately 0.001% to approximately 50%, approximately 0.001% toapproximately 40%, approximately 0.01% to approximately 30%,approximately 0.02% to approximately 29%, approximately 0.03% toapproximately 28%, approximately 0.04% to approximately 27%,approximately 0.05% to approximately 26%, approximately 0.06% toapproximately 25%, approximately 0.07% to approximately 24%,approximately 0.08% to approximately 23%, approximately 0.09% toapproximately 22%, approximately 0.1% to approximately 21%,approximately 0.2% to approximately 20%, approximately 0.3% toapproximately 19%, approximately 0.4% to approximately 18%,approximately 0.5% to approximately 17%, approximately 0.6% toapproximately 16%, approximately 0.7% to approximately 15%,approximately 0.8% to approximately 14%, approximately 0.9% toapproximately 12%, approximately 1% to approximately 10% w/w, w/v orv/v. It is understood that the final concentration is dependent on manyfactors known to persons skilled in the art including, but not limitedto, location and size of the target site.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g,0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g,0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g,0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g of the one or morepharmacologically active detergents herein.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of more than 0.0001 g, 0.0002 g, 0.0003 g,0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g,0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g,0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g,0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g,0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g,5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g of theone or more pharmacologically active detergents herein.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of 0.0001-10 g, 0.0005-9 g, 0.001-8 g,0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g of the one ormore pharmacologically active detergents herein.

In any of the embodiment herein, a composition can comprise, consistessentially of, or consist of at least 2, 3, 4, 5, 6, 7, 8, 9, or 10detergents.

In any of the embodiments herein, a composition can include one or morephospholipids (e.g., phosphatidylcholine). Preferably, the amount ofphospholipids in a composition/unit dose herein is at a concentrationless than 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,1%, 0.95%, 0.9%, 0.85%, 0.8%, 0.75%, 0.7%, 0.65%, 0.6%, 0.55%, 0.5%,0.45%, 0.4%, 0.35%, 0.3%, 0.25%, 0.2%, 0.15%, 0.1%, 0.09%, 0.08%, 0.07%,0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v, or v/v of the composition or unit dose. In preferred embodiments,the amount of phospholipids in a composition is at a concentration lessthan 5% w/w, w/v, or v/v.

In one embodiment of the present invention, a medical composition forthe non-surgical reduction of localized fat deposits in a patient isprovided which comprises at least one pharmacologically activedetergent, optionally at least one pharmaceutically acceptable excipientand optionally at least one additional active ingredient wherein themedical composition and contains less than 20%, 19%, 18%, 17%, 16%,15%,14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%,0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%,0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%,0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%,0.0003%, 0.0002%, or 0.0001% w/w, w/v, or v/v of phospholipids (such asphosphatidylcholine), or more preferably does not contain anyphospholipids, such as phosphatidylcholine. The term “less than” as usedherein when refers generally to a composition containing somephosphatidylcholine, but in some embodiments refers to 0%phosphatidylcholine.

In an embodiment of the present invention, the pharmacologically activedetergent composition contains at least one pharmacologically activedetergent, optionally at least one pharmaceutically acceptable excipientand optionally at least one additional active ingredient, and whereinthe pharmacologically active detergent composition contains less than20%, 19%, 18%, 17%, 16%, 15%,14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%,5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%,0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v, or v/v of phospholipids (such as phosphatidylcholine), or morepreferably does not contain any phospholipids, such asphosphatidylcholine.

In embodiments of the present invention, the pharmacologically activedetergent composition is administered by subcutaneous injection directlyinto fat tissue.

In an embodiment of the present invention, the localized fataccumulation is lower eyelid fat herniation, lipomas, lipodystrophy,buffalo hump lipodystrophy or fat deposits associated with cellulite.Localized fat accumulations can be present in, for example, under eye,under chin, under arm, buttock, calf, ankle, back, thigh, or stomach.Thus, the present invention contemplates treatment of adipose tissuedisorders such as lipomas, Dercum's disease, Madelung's neck, lipedema,piezogenic nodules, xanthelasma, lipodystrophy, and cellulite.

In another embodiment of the present invention, a medical composition isprovided for reducing localized accumulation of fat in a patient withlower eyelid fat herniation comprising a fat solubilizing amount ofdeoxycholic acid, and the medical composition contains less than 20%,19%, 18%, 17%, 16%, 15%,14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%,0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%,0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%,0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w, w/v, or v/vphospholipids (such as phosphatidylcholine), or more preferably does notcontain any phospholipids, such as phosphatidylcholine.

In an embodiment of the present invention a non-liposuction method forthe non-surgical reduction of localized fat deposits in a patient isprovided comprising the non-surgical administration of apharmacologically active detergent composition consisting essentially ofat least one pharmacologically active detergent, optionally at least onepharmaceutically acceptable excipient and optionally at least oneadditional active ingredient, and the medical composition and preferablycontains less than 20%, 19%, 18%, 17%, 16%, 15%,14%, 13%, 12%, 11%, 10%,9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%,0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%,0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%,0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or0.0001% w/w, w/v, or v/v phospholipids (such as phosphatidylcholine), ormore preferably does not contain any phospholipids, such asphosphatidylcholine.

In any of the compositions herein the ratio between the detergent(s) andphospholipids is such there is more detergents by mass thanphospholipids. For example, the mass ration of detergent(s) andphospholipids may be 1:0.5, 1:0.05, 1.0.005, etc. In some embodiments,the concentration of the phospholipids (e.g., phosphatidylcholine) in%w/v is less than the concentration of % w/v of the detergent(s). Forexample, a composition may have 5% w/v sodium deoxycholate and 4% w/vphosphatidylcholine.

Unit Dose

The present invention also contemplates a unit dose of the compositionsherein. Such unit dose can have, for example, a total volume of lessthan 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL,50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6, mL 5 mL, 4 mL, 3mL, 2 mL, 1 mL, 0.9 mL, 0.8 mL, 0.7 mL, 0.6 mL, 0.5 mL, 0.4 mL, 0.3 mL,0.2 mL, 0.1 mL, 0.09 mL, 0.08 mL, 0.07 mL, 0.06 mL, 0.05 mL, 0.04 mL,0.03 mL, 0.02 mL, 0.01 mL, 0.009 mL, 0.008 mL, 0.007 mL, 0.006 mL, 0.005mL, 0.004 mL, 0.003 mL, 0.002 mL, 0.001 mL, 0.0009 mL, 0.0008 mL, 0.0007mL, 0.0006 mL, 0.0005 mL, 0.0004 mL, 0.0003 mL, 0.0002 mL, or 0.0001 mL.In some embodiments, such unit dose has a total volume of more than 0.2mL and less than 500 mL. In some embodiments, such unit dose has a totalvolume of less than 0.1 mL. In some embodiments, such unit dose has atotal volume of less than 0.1 mL. In some embodiments, such unit dosehas total volume of 0.1-0.2 mL (inclusive of 0.1 mL and 0.2 mL). In someembodiments, such unit dose has total volume of less than 0.1 andgreater than 0.2.

In some embodiments, the present invention contemplates administration acomposition or unit dose that is greater 0.0001 mL, 0.0005 mL, 0.001 mL,0.005 mL, 0.01 mL, 0.05 mL, 0.1 mL, 0.5 mL, 1 mL, 5 mL, 10 mL, 50 mL,100 mL of total volume to target site.

In some embodiments, the present invention contemplates administrationof a unit dose having a total volume in the range of 0.0001-500 mL,0.0005-400 mL, 0.001-300 mL, 0.005-200 mL, 0.01-100 mL, 0.05-90 mL,0.06-80 mL, 0.07-70 mL, 0.08-60 mL, 0.09-50 mL, 0.1-40 mL, 0.2-30 mL,0.3-29 mL, 0.4-28 mL, 0.5-27 mL, 0.6-26 mL, 0.7-25 mL, 0.8-24 mL, 0.9-23mL, 10-22 mL, 11-21 mL, 12-20 mL, 13-19 mL, 14-18 mL, or 15-17 mL pertarget site.

Other embodiments contemplate administration a total volume of acomposition that is in the range of 0.01-30 mL, 0.02-20 mL, 0.03-10 mLof total volume of a composition per target site. Other embodimentscontemplate administration of 0.2-500 mL of total solution to a targetsite, 0.1-0.2 mL total solution to a target site, less than 0.1 mL(optionally excluding 0.03 mL and 0.05 mL per target site).

A unit dose can comprises, consists essentially of, or consists of anamount of the one or more pharmacologically active detergents asdisclosed in the compositions herein. A unit dose can further includephospholipids such as phosphatidylcholine at concentrations and unitsidentified in the composition section above. For example, a preferredunit dose has less than 5 g of pharmacologically active detergent(s)and/or less than 5% phospholipids, such as phosphatidylcholine.

In preferred embodiments, a unit dose comprises, consists essentiallyof, or consists of one or more pharmacologically active detergent(s) inan inejctable formulation wherein the unit dose has a total volume ofless than 500 mL, but more than 0.2 mL. Such unit dose may have lessthan 5%, w/w, w/v or v/v phospholipids (e.g., phosphatidylcholine).

In some embodiments, a unit dose comprises of more than 0.1% w/w, w/v orv/v of the one or more detergents herein and the unit dose has a totalvolume of more than 0.2 mL and less than 500 mL. In some embodiments, aunit dose comprises of more than 0.1% w/w, w/v or v/v of the one or moredetergents herein and the unit dose has a total volume of less than 0.1mL, optionally excluding 0.03 mL and 0.05 mL.

In some embodiments, a unit dose comprises less than 0.01 g of the oneor more detergents and has a total volume of less than 500 mL.

For example, in some embodiments, a unit dose comprises of less than0.1% or 0.01% by weight of the one or more detergents herein.

In some embodiments, a unit dose has less than 0.9% w/w or more than 13%w/w of the one or more detergents herein and has a total volume of0.1-0.2 mL.

The unit dose will depend, in part, on the target area, amount of fat,and desired result.

Salts and Esters

The present invention also contemplates pharmaceutically acceptablesalts and esters of the detergents herein. Such salts and esters aremeant to be those salts and esters which are within the scope of soundmedical judgment, suitable for use in contact with the tissues of humansand animals without undue toxicity, irritation, allergic response, andthe like, commensurate with a reasonable benefit/risk ratio, andeffective for their intended use.

Among the more common pharmaceutically acceptable salts and esters arethe acetate, estolate (lauryl sulfate salt of the propionate ester),ethyl succinate, gluceptate (glucoheptonate), lactobionate, stearate,and hydrochloride forms. Other acid salts contemplated herein are thefollowing: adipate, alginate, aspartate, benzoate, benzene sulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,fumarate, gluconate, glycerophosphate, hemisulfate, heptonate,hexanoate, hydrobromide, hydroiodide, 2-hydroxy ethanesulfonate,lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,oxalate, pamoate, pantothenate, pectinate, persulfate,3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate,thiocyanate, tosylate, and undecanoate.

Basic nitrogen-containing groups can be quaternized with such agents aslower alkyl halides, such as methyl, ethyl, propyl and butyl chloride,bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl,and diamyl sulfates; long chain halides such as decyl, lauryl, myristyland stearyl chlorides, bromides and iodides; aralkyl halides like benzyland phenethyl bromides and others.

In preferred embodiments, one or more of the detergents herein are bilesalts. Bile salts herein may be formed with inorganic bases, ammonia,organic bases, inorganic acids, organic acids, basic amino acids,halogen ions or the like, and inner salts. Examples of the inorganicbase include alkali metal (e.g., Na and K) and alkaline earth metal(e.g., Mg). Examples of the organic base include procaine,2-phenylethylbenzylamine, dibenzylethylenediamine, etanolamine, dietanolamine, tris(hydroxymethyl)aminomethane, polyhydroxyalkylamine, andN-methyl glucosamine. Examples of the inorganic acid includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, andphosphoric acid. Examples of the organic acid include p-toluene sulfonicacid, methanesulfonic acid, formic acid, trifluoroacetc acid and maleicacid. Examples of the basic amino acid include lysine, arginine,ornithine and histidine.

Bile acids may be present as their esters, for example, but not limitedto, optionally substituted C1-C6 alkyl, C2-C6 alkenyl, C3-C10cycloalkyl, C3-C10 cycloalkyl(C1-C6)alkyl, optionally substituted C6-C10aryl, optionally substituted C7-C12 aralkyl, di(C6-C10)arylmethyl,tri(C6-C10)arylmethyl, and substituted silyl.

Examples of the optionally substituted C1-6 alkyl include e.g., methyl,ethyl, n-propyl, n-butyl, t-butyl, n-pentyl, and n-hexyl, each may besubstituted with benzyloxy, C1-4 alkylsulfonyl (e.g., methanesulfonyl),trimethylsilyl, halogen (e.g., F, Cl, and Br), acetyl, nitrobenzoyl,mesylbenzoyl, phthalimide, succinoylimide, benzenesulfonyl,phenylthio,di-C1-4alkylamino (e.g., dimethylamino), pyridyl, C1-4alkylsulfinyl(e.g., methanesulfinyl), cyano and the like. Such substituted C1-6 alkylinclude e.g., benzyloxymethyl, 2-methanesulfonylethyl,2-trimethylsilylethyl, 2,2,2-trichloroethyl, 2-iodoethyl, acetylmethyl,p-nitrobenzoylmethyl, p-mesylbenzoylmethyl, phthalimidemethyl,succinoylimidemethyl, benzenesulfonylmethyl, phenylthiomethyl, and1-dimethylaminoethyl. The above C2-6 alkenyl includes e.g., vinyl, aryl,1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, 1,1-dimethylaryl,3-methyl and 3-butenyl. The above C3-10 cycloalkyl includes e.g.,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,norbornyl, and adamantyl. The above C3-10 cycloalkyl(C1-6)alkyl includese.g., cyclopropylmethyl, cyclopentylmethyl, and cyclohexylmethyl. Theabove C6-10 aryl includes e.g., phenyl, .alpha.-naphthyl, 8-naphthyl,and biphenyl, each may be substituted with nitro, halogen (e.g., F, Cl,and Br) or the like, and such substituted aryl includes e.g.,p-nitrophenyl and p-chlorophenyl. The above optionally substituted C7-12aralkyl includes e.g., benzyl, 1-phenylethyl, 2-phenylethyl,phenylpropyl and naphthylmethyl, each may be substituted with nitro,C1-4 alkoxy (e.g., methoxy), C1-4 alkyl (e.g., methyl, ethyl), hydroxyor the like. Such substituted group is exemplified by p-nitrobenzyl,p-methoxybenzyl (PMB), or 3,5-di-t-butyl-4-hydroxybenzyl. The abovedi(C6-10 aryl)methyl includes benzhydryl and the C6-10 arylmethylincludes trityl, and the substituted silyl includes trimethylsilyl andtert-butyldimethylsilyl, Examples of the active ester include organicphosphate esters (e.g., diethoxy phosphate ester and diphenoxy phosphateester), cyanomethyl ester, and the active thioester includes estersformed with aromatic heterocyclicthio compound (e.g., 2-pyridilthioester).

Examples, of other reactive derivative of bile acids include acidhalides, acid azides, acid anhydrides, mixed acid anhydride, activeamide, and active thioester. The acid halide includes acid chloride andacid bromide; the mixed acid anhydride includes mixedmonoalkylcarboxylic acid anhydride, mixed alphatic carboxylic acidanhydride, aromatic carboxylic acid anhydride, organic sulfonic acidanhydride, the active amide includes amide formed with heterocycliccompound containing N atom, for example.

Micelles

Detergents, including bile acids, are micelle-forming compounds. It isbelieved that the presence of the micelles significantly increases thesolubility of hydrophobic molecules not ordinarily soluble in water(e.g., the lipids that comprise cell membranes) by burying theirhydrophobic portions away from aqueous solvent (e.g., water). As will beappreciated by those skilled in the art, a micelle is a colloidalaggregate of amphipathic molecules in which the polar hydrophilicportions of the molecule extend outwardly while the non-polarhydrophobic portions extend inwardly.

In some embodiments, the present invention contemplates homogenousmicelles (micelles produced by a single detergent), while in otherembodiments, the present invention contemplates mixed micellarformations (micelles produced by two or more compounds—one of which is adetergent).

In some embodiments, an average particle size of micelles in acomposition of the present invention is contemplated to be in the rangeof 1 nanometer to 100 micrometers, 10 nanometers to 50 micrometers, 100nanometers to 1 micrometers, etc. Moreover, the shape of the micelle canvary and can be, for example, prolate, oblate or spherical; sphericalmicelles are most typical.

In some embodiments, at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 55%, 60%, 65%, 70%, 75%, 80% 85%, 90%, 95%, 99% of the detergent inthe compositions herein is in micellar formation. In other embodiments,less than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or 5% of thedetergent in the compositions herein is in micellar formation. In otherembodiments, about 10-90%, 20-80%, 30-70%, 40-60%, or about 50% of thedetergent of the compositions herein is in micellar formation.

In some embodiments, an average size of a micelle in a composition ofthe present invention may be less than 10⁻⁵, 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹. Insome embodiments, an average size of a micelle in a composition of thepresent invention may be greater than 10⁻⁵, 10⁻⁸, 10⁻⁷, 10⁻⁸, 10⁻⁹. Insome embodiments, an average size of a micelle in a composition of thepresent invention may be in the range of 1×10⁻⁵ to 9×10⁻⁵; 1×10⁻⁶ to9×10⁻⁸; 1×10⁻⁷ to 9×10⁻⁷;1×10⁻⁸ to 9×10⁻⁸; 1×10⁻⁹ to 9×10⁻⁹.

In some embodiments, an average molecular weight of a micelle in acomposition of the present invention may be less than 100,000 Daltons,50,000 Daltons, 40,000 Daltons, 30,000 Daltons, 20,000 Daltons, 10,000Daltons, 9,000 Daltons, 8,000 Daltons, 7,000 Daltons, 6,000 Daltons,5,000 Daltons, 4,000 Daltons, 3,000 Daltons, 2,000 Daltons, 1,000Daltons, or 500 Daltons. In some embodiments, an average molecularweight of a micelle in a composition of the present invention may begreater than 500 Daltons, 1,000 Daltons, 1,500 Daltons, 2,000 Daltons,2,500 Daltons, 3,000 Daltons, 3,500 Daltons, 4,000 Daltons, 4,500Daltons, 5,000 Daltons, 5,500 Daltons, 6,000 Daltons, 6,500 Daltons,7,000 Daltons, 7,500 Daltons, 8,000 Daltons, 8,500 Daltons, 9,000Daltons, 9,500 Daltons, 10,000 Daltons, or 15,000 Daltons. In someembodiments, an average molecular weight of a micelle in a compositionof the present invention may be in the range of 100-20,000 Daltons,1,000-10,000 Daltons, 2,000-1,000 Daltons, or 3,000-5,000 Daltons.

Second Therapeutic Agents

In yet another embodiment of the present invention the compositionsherein can be co-formulated, co-administered, and/or co-marketed with asecond therapeutic agent.

Non-limiting examples of second therapeutic agents include:anti-microbial agents, vasoconstrictors, anti-thrombotic agents,anti-coagulation agents, suds-depressants, anti-inflammatory agents,analgesics, dispersion agents, anti-dispersion agents, penetrationenhancers, steroids, tranquilizers, muscle relaxants, and anti-diarrheaagents.

Anti microbial agents suitable for use with the compositions, methods,and kits herein include, but not limited to, anti-bactericidal agents,anti-fungal agents, anti-viral agents or the like, and are preferablyefficacious against a broad spectrum of microbes.

Examples of anti-bacterial agents include, but not limited to,benzalkonium chloride, benzoic acid, benzoxonium chloride, benzylalcohol, 2-bromo-2-nitropropane-1,3-diol, 5-bromo-5-nitro-1,3-dioxane,bromochlorophene, camphor benzalkonium methosulfate, captan, cetrimoniumbromide, cetrimonium chloride, cetylpyridinium chloride, climbazol,chloracetamide, chiorhexidine and its salts, p-chloro-m-cresol,chlorphenesin, chloroxylenol, chlorophen, chlorobutanol, o-cymen-5-ol,dehydroacetic acid, dibromodicyanobutan, dibromohexamidin,dibromopropamidin, dichlorobenzyl alcohol, dichlorophenylimidazoldioxolan, dimethyloxazolidin, DMDM hydantoin, dodecylguanidineacetate, hexamidine diisothionate, hexachlorophen, hexetidin,iodopropynyl butylcarbamate, lauryl isoquinolinium bromide,methyldibromo glutaronitrile, methylolchloracetamide, phenethyl alcohol,phenoxyethanol, phenoxypropanol, o-phenylphenol, piroctone olamine,polyaminopropyl biguanide, potassium sorbate, potassium undecylenoylhydrolyzed collagen, quaternium-15, salicylic acid, sodium benzoate,sodium dehydroacetate, sodium hydroxymethylglycinate, sodiumo-phenylphenate, sorbic acid, triclocarban, triclosan, undecylenic acidand its derivatives, zinc cysteate, zinc gluconate, zinc pyrithione, orzinc sulfate. Derivatives of undecylenic acid useful as anti-microbialagents are e.g. esters, such as methyl ester, isopropyl ester, glycerylester, ethoxylated soya sterol ester, or ethoxylated PHB ester, oramides, such as monoethanolamide, monoethanolamide derivatives such asmonoethanolamide (MEA) sulfosuccinate salts, diethanolamide, proteincondensates, e.g. potassium undecylenoyl hydrolyzed animal collagen, andquaternized 3-aminopropyl-amide, e.g. undecylenamidopropyltrimoniummethosulfate. Specific examples of suitable fungicidal/fungistaticagents include, without limitation, dithiocarbamates, phthalimides,dicarboximides, organophosphates, benzimidazoles, carboxanilides,phenylamides, phosphites, and the like.

Other examples of anti-bacterial agents include, but are not limited to,erythromycin, clarithromycin, penicillins, cephalosporins,aminoglycosides, sulfonamides, macrolides, tetracyclins, lincosides,quinolones, chloramphenicol, vancomycin, metronidazole, rifampin,isoniazid, spectinomycin, trimethoprim, sulfamethoxazole, penems,carbapenems, monobactams mupirocin, neomycin sulfate bacitracin,polymyxin B, 1-ofloxacin, tetracyclines (chlortetracyclinehydrochloride, oxytetracycline hydrochloride and tetrachcyclinehydrochoride), clindamycin phsphate, gentamicin sulfate, benzalkoniumchloride, benzethonium chloride, hexylresorcinol, methylbenzethoniumchloride, phenol, quaternary ammonium compounds, triclocarbon,triclosan, tea tree oil, and their pharmaceutically acceptable salts.and the pharmaceutically acceptable salts and esters thereof.

Other examples of anti-bacterial agents include, but are not limited to,Acrofloxacin, Amoxicillin plus clavulonic acid (i.e. Augmentin),Amikacin, Amplicillin, Apalcillin, Apramycin, Astromicin, Arbekacin,Aspoxicillin, Azidozillin, Azithromycin, Azlocillin, Bacitracin,Benzathine penicillin, Benzylpenicillin, Carbencillin, Cefaclor,Cefadroxil, Cefalexin, Cefamandole, Cefaparin, Cefatrizine, Cefazolin,Cefbuperazone, Cefcapene, Cefdinir, Cefditoren, Cefepime, Cefetamet,Cefixime, Cefmetazole, Cefminox, Cefoperazone, Ceforanide, Cefotaxime,Cefotetan, Cefotiam, Cefoxitin, Cefpimizole, Cefpiramide, Cefpodoxime,Cefprozil, Cefradine, Cefroxadine, Cefsulodin, Ceftazidime, Ceftriaxone,Cefuroxime, Chlorampenicol, Chlortetracycline, Ciclacillin, Cinoxacin,Ciprofloxacin, Clarithromycin, Clemizole penicillin, Clindamycin,Cloxacillin, Daptomycin, Demeclocycline, Desquinolone, Dibekacin,Dicloxacillin, Dirithromycin, Doxycycline, Enoxacin, Epicillin,Erthromycin, Ethambutol, Fleroxacin, Flomoxef, Flucloxacillin,Flumequine, Flurithromycin, Fosfomycin, Fosmidomycin, Fusidic acid,Gatifloxacin, Gemifloxaxin, Gentamicin, Imipenem, Imipenem plusCilistatin combination, Isepamicin, Isoniazid, Josamycin, Kanamycin,Kasugamycin, Kitasamycin, Latamoxef, Levofloxacin, Lincomycin,Linezolid, Lomefloxacin, Loracarbaf, Lymecycline, Mecillinam, Meropenem,Methacycline, Methicillin, Metronidazole, Mezlocillin, Midecamycin,Minocycline, Miokamycin, Moxifloxacin, Nafcillin, Nafcillin, Nalidixicacid, Neomycin, Netilmicin, Norfloxacin, Novobiocin, Oflaxacin,Oleandomycin, Oxacillin, Oxolinic acid, Oxytetracycline, Paromycin,Pazufloxacin, Pefloxacin, Penicillin G, Penicillin V, Phenethicillin,Phenoxymethyl penicillin, Pipemidic acid, Piperacillin, Piperacillin andTazobactam combination, Piromidic acid, Procaine penicillin,Propicillin, Pyrimethamine, Rifabutin, Rifamide, Rifampicin, RifamycinSV, Rifapentene, Rokitamycin, Rolitetracycline, Roxithromycin,Rufloxacin, Sitafloxacin, Sparfloxacin, Spectinomycin, Spiramycin,Sulfadiazine, Sulfadoxine, Sulfamethoxazole, Sisomicin, Streptomycin,Sulfamethoxazole, Sulfisoxazole, Synercid (Quinupristan-Dalfopristancombination), Teicoplanin, Telithromycin, Temocillin, Tetracycline,Tetroxoprim, Thiamphenicol, Ticarcillin, Tigecycline, Tobramycin,Tosufloxacin, Trimethoprim, Trimetrexate, Trovafloxacin, Vancomycin, andVerdamicin.

Vasoconstrictor agents suitable for use with the compositions of thepresent invention can include, for example, dihydroergotamine,ergotamine and methysergide, pharmaceutically-acceptable salts thereof,

Anti-thrombotic agents suitable for use with the compositions of thepresent invention can include, for example, argatroban, iloprost,lamifiban, taprostene, tirofiban, tissue plasminogen activator (naturalor recombinant), tenecteplase (TNK), and lanoteplase (nPA); factor VIIainhibitors; factor Xa inhibitors; thrombin inhibitors (such as hirudinand argatroban); PAI-1 inhibitors (i.e., inactivators of tissueplasminogen activator inhibitors); alpha2-antiplasmin inhibitors;streptokinase, urokinase and prourokinase; and anisoylated plasminogenstreptokinase activator complex. anti-coagulants (e.g. hirudin, heparin,etc.), plasminogen activators (e.g. t-PA, urokinase, etc.), fibrinolyticenzymes (e.g. plasmin, subtilisin, etc.), anti-platelet-aggregationagents (e.g. prostacyclin, aspirin, etc.) and the like.

Anti-coagulation agents suitable for use with the compositions of thepresent invention can include, for example, cilostazol (PLETAL®,Otsuka), clopidogrel (PLAVIX®, Sanofi), ticlopidine (TICLID®, Syntex),tirofiban (AGGRASTAT®, Merck), eptifibatide (INTEGRILIN®, CORTherapeutics), abciximab (REOPRO®, Eli Lill y), anagrelide (AGRYLIN®,Roberts), dipyridamole (PERSANTIN®, Boehringer Ingelheim), aspirin(ECOTR®, and others), dipyridamole/aspirin (AGGRENOX®, BoehringerIngelheim), dalteparin (FRAGMIN®, Pharmacia), enoxaparin (LOVENOX®,Aventis), tinzaparin (INNOHE®, DuPont), heparin (various), danaparoid(ORGANON®, Organon), antithrombin III (THROMBATE®, Bayer), lepirudin(REFLUDAN®, Hoechst-Marion Roussel), argatroban (ACOVA®,SmithKlineBeecham), bivalirudin (ANGIOMAX®, Medicines Company), warfarin(COUMADIN®, DuPont) anisidione (MIRADON®, Schering), alteplase(ACTIVASE®, Genetech), reteplase (RETAVASE®, Boehringer Mannheim),tenecteplase (TNKASE®, Genentech), drotrecogin (XIGRIS®, Eli Lilly),anistreplase (EMINASE®, Roberts), streptokinase (STREPTASE®, Astra),urokinase (ABBOKINASE®, Abbott) and combinations thereof.

Suds-depressants suitable for use with the compositions, methods andkits of the present invention can include, for example, monocarboxylicfatty acid and soluble salts thereof. The monocarboxylic fatty acids andsalts thereof used as suds suppressor may have hydrocarbyl chains of 1to about 50 carbon atoms, about 10 to about 24 carbon atoms, or about 12to about 18 carbon atoms. Suitable salts include the alkali metal saltssuch as sodium, potassium, and lithium salts, and ammonium andalkanolammonium salts. Additional suds-depressnats include, for example,high molecular weight hydrocarbons such as paraffin, fatty acid esters(e.g., fatty acid triglycerides), fatty acid esters of monovalentalcohols, aliphatic C₁₈-C₄₀ ketones (e.g., stearone), etc. Othersuds-depressants include N-alkylated amino triazines such as tri- tohexa-alkylmelamines or di- to tetra-alkyldiamine chlortriazines formedas products of cyanuric chloride with two or three moles of a primary orsecondary amine containing 1 to 24 carbon atoms, propylene oxide, andmonostearyl phosphates such as monostearyl alcohol phosphate ester andmonostearyl di-alkali metal (e.g., K, Na, and Li) phosphates andphosphate esters. The hydrocarbons such as paraffin and haloparaffin canbe utilized in liquid form. It is also known to utilize waxyhydrocarbons, preferably having a melting point below about 100° C. Thehydrocarbons constitute a preferred category of suds suppressor fordetergent compositions. The hydrocarbons, thus, include aliphatic,alicyclic, aromatic, and heterocyclic saturated or unsaturatedhydrocarbons having from about 12 to about 70 carbon atoms. The term“paraffin,” as used in this suds suppressor discussion, is intended toinclude mixtures of true paraffins and cyclic hydrocarbons.

Another example of suds suppressors comprises silicone suds suppressors.This category includes the use of polyorganosiloxane oils, such aspolydimethylsiloxane, dispersions or emulsions of polyorganosiloxaneoils or resins, and combinations of polyorganosiloxane with silicaparticles wherein the polyorganosiloxane is chemisorbed or fused ontothe silica. Examples also include , but not limited to, silicones, andsilica-silicone mixtures. Silicones can be generally represented byalkylated polysiloxane materials while silica is normally used in finelydivided forms exemplified by silica aerogels and xerogels andhydrophobic silicas of various types. Silicone suds controlling agent,DC-544, is commercially available from Dow Corning, which is asiloxane-glycol copolymer. Other preferred suds controlling agent arethe suds suppressor system comprising a mixture of silicone oils and2-alkyl-alcanols. Suitable 2-alkyl-alkanols are 2-butyl-octanol whichare commercially available under the trade name Isofol 12™ andsilicone/silica mixture in combination with fumed nonporous silica suchas Aerosil™.

Examples of anti-dispersion agents include, but are not limited to,sucrose, glyercerol, and glycerin.

Steroids suitable for use with the compositions of the present inventioncan include, for example, betamethasone, chloroprednisone, clocortolone,cortisone, desonide, dexamethasone, desoximetasone, difluprednate,estradiol, fludrocortisone, flumethasone, flunisolide, fluocortolone,fluprednisolone, hydrocortisone, meprednisone, methylprednisolone,paramethasone, prednisolone, prednisone, pregnan-3-alpha-ol-20-one,testosterone, and triamcinolone. estradiol, estron, estriol,polyestradiol, polyestriol, dienestrol, diethylstilbestrol,dihydroergosterone, cyproterone, danazol, testosterone, progesterone,norethindrone, levonorgestrol, ethynodiol, norgestimate, gestanin,3-keton-desogestrel, demegestone, promethoestrol, testosterone,spironolactone, and esters thereof, budesonide, rofleponide, rofleponidepalmitate, ciclesonide, momethasone furoate, fluticasone propionate,tipredane, fluocinolone acetonide, flunisolide, flumethasone,dexamethasone, beclomethasone dipropionate, deflazacort, cortivazol, orcortisol and/or hydrocortisol, prednisone, fluorometholone acetate,dexamethasone sodium phosphate, suprofen, fluorometholone, andmedrysone, optionally in their pure isomeric forms (where such formsexist) and in the forms of their pharmaceutically acceptable salts.

Anti-inflammatory agents suitable for use with the compositions of thepresent invention can include both steroidal anti-inflammatory agentsand non-steroidal anti-inflammatory agents. Suitable steroidalanti-inflammatory agent can include, although are not limited to,corticosteroids such as hydrocortisone, hydroxyltriamcinolonealphamethyl dexamethasone, dexamethasone-phosphate, beclomethasonedipropionate, clobetasol valerate, desonide, desoxymethasone,desoxycorticosterone acetate, dexamethasone, dichlorisone, diflorasonediacetate, diflucortolone valerate, fluadrenolone, fluclaroloneacetonide, fludrocortisone, flumethasone pivalate, fluosinoloneacetonide, fluocinonide, flucortine butylester, fluocortolone,fluprednidene (fluprednylidene)acetate, flurandrenolone, halcinonide,hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone,triamcinolone acetonide, cortisone, cortodoxone, flucetonide,fludrocortisone, difluorosone diacetate, fluradrenalone acetonide,medrysone, amciafel, amcinafide, betamethasone and the balance of itsesters, chlorprednisone, chlorprednisone acetate, clocortelone,clescinolone, dichlorisone, difluprednate, flucloronide, flunisolide,fluoromethalone, fluperolone, fluprednisolone, hydrocortisone valerate,hydrocortisone cyclopentylproprionate, hydrocortamate, meprednisone,paramethasone, prednisolone, prednisone, beclomethasone dipropionate,betamethasone dipropionate, triamcinolone, and mixtures thereof can beused.

A second class of anti-inflammatory agents which is useful in thecompositions of the present invention includes the nonsteroidalanti-inflammatory agents. A variety of compounds encompassed by thisgroup are well-known to those skilled in the art. Suitable non-steroidalanti-inflammatory agents useful in the compositions of the presentinvention include, but are not limited to: the oxicams, such aspiroxicam, isoxicam, tonexicam, sudoxicam, and CP-14,304; thesalicylates, such as salicylic acid, aspirin, disalcid, benorylate,trilisate, safapryn, solprin, diflunisal, and fendosal; the acetic acidderivatives, such as diclofenac, fenclofenac, indomethacin, sulindac,tolmetin, isoxepac, furofenac, tiopinac, zidometacin, acematacin,fentiazac, zomepiract, clidanac, oxepinac, and felbinac; the fenamates,such as mefenamic, meclofenamic, flufenamic, niflumic, and tolfenamicacids; the propionic acid derivates, such as ibuprofen, naproxen,benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen,indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen,tioxaprofen, suprofen, alminoprofen, and tiaprofenic; and the pyrazoles,such as phenybutazone, oxyphenbutazone, feprazone, azapropazone, andtrimethazone. Mixtures of these nonsteroidal anti-inflammatory agentscan also be employed, as well as the pharmaceutically-acceptable saltsand esters of these agents.

Analgesics suitable for use with the pharmacologically active detergentcomposition of the present invention to reduce discomfort due toinflammation after subcutaneous injection of the formulation of thepresent invention include, but are not limited to, injectable localamine and ester anesthetics. Non-limiting examples of analgesics includelidocaine, mepivacaine, bupivacaine, procaine, chloroprocaine,etidocaine, prilocaine dyclonine, hexylcaine, procaine, cocaine,ketamine, pramoxine, propophol, phenol and tetracaine. Mixtures of theseanalgesics can also be employed, as well as the pharmaceuticallyacceptable salts and esters or these agents. Other examples ofanalgesics include opioids. Examples of opioids include morphine, or asalt thereof, such as the sulphate, chloride, or hydrochloride. Other1,4-hydroxymorphinan opioid analgesics that may be used herein includethose such as naloxone, meperidine, butorphanol or pentazocine, ormorphine-6-glucuronide, codeine, dihydrocodeine, diamorphine,dextropropoxyphene, pethidine, fentanyl, alfentanil, alphaprodine,buprenorphine, dextromoramide, diphenoxylate, dipipanone, heroin(diacetylmorphine), hydrocodone (dihydrocodeinone), hydromorphone(dihydromorphinone), levorphanol, meptazinol, methadone, metopon(methyldihydromorphinone), nalbuphine, oxycodone(dihydrohydroxycodeinone), oxymorphone (dihydrohydroxymorphinone),phenadoxone, phenazocine, remifentanil, tramadol, or a salt of any ofthese. The opioid used in the method of the invention may comprise anycombination of the aforementioned compounds. Naloxone is also includedwithin the definition of an opioid. Especially preferred analgesicswhich may be use include hydromorphone, oxycodone, morphine, e.g.morphine sulphate and fentanyl and/or pharmaceutically-acceptable saltsthereof.

Suitable tranquilizer and sedative drugs that may included in the kitsor compositions of the present invention include chlordiazepoxide,benactyzine, benzquinamide, flurazepam, hydroxyzine, loxapine,promazine, and/or acceptable salts and esters thereof.

Suitable muscle relaxant drugs that may be included in the kits orcompositions of the present invention include cinnamedrine,cyclobenzaprine, flavoxate, orphenadrine, papaverine, mebeverine,idaverine, ritodrine, dephenoxylate, dantrolene, azumolene, and/orpharmaceutically-acceptable salts thereof.

Suitable anti-diarrhea drugs may be included in the kits or compositionsof the present invention include, for example, loperamide, and/orpharmaceutically-acceptable salts thereof.

Second therapeutic agents may be co-formulated and/or co-administeredwith the one or more pharmacologically active detergents herein. In suchco-formulations, a second therapeutic agent may be at a concentration ofless than 20%, 19%, 18%, 17%, 16%, 15%,14%, 13%, 12%, 11%, 10%, 9%, 8%,7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%,0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v or v/v.

In some embodiments, a second therapeutic agent may be co-formulatedwith the one or more pharmacologically active detergents herein. In suchco-formulation, the second therapeutic agent may be at a concentrationgreater than 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25%18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%,15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25%13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%,10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%,7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%,4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%,2%, 1.75%, 1.50%, 125% , 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%,0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%,0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%,0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w/w,w/v or v/v.

In some embodiments, a second therapeutic agent may be co-formulatedwith the one or more pharmacologically active detergents herein suchthat the final formulation has a concentration of the second therapeuticagent that is in the range of from approximately 0.001% to approximately50%, approximately 0.001% to approximately 40%, approximately 0.01% toapproximately 30%, approximately 0.02% to approximately 29%,approximately 0.03% to approximately 28%, approximately 0.04% toapproximately 27%, approximately 0.05% to approximately 26%,approximately 0.06% to approximately 25%, approximately 0.07% toapproximately 24%, approximately 0.08% to approximately 23%,approximately 0.09% to approximately 22%, approximately 0.1% toapproximately 21%, approximately 0.2% to approximately 20%,approximately 0.3% to approximately 19%, approximately 0.4% toapproximately 18%, approximately 0.5% to approximately 17%,approximately 0.6% to approximately 16%, approximately 0.7% toapproximately 15%, approximately 0.8% to approximately 14%,approximately 0.9% to approximately 12%, approximately 1% toapproximately 10% w/w, w/v or v/v. It is understood that the finalconcentration is dependent on many factors known to persons skilled inthe art including, but not limited to, location and size of thetreatment site.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g,2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g,0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g,0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g,0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g of the one or moresecond therapeutic agents herein.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of more than 0.0001 g, 0.0002 g, 0.0003 g,0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g,0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g,0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g,0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g,0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5, 3 g, 3.5, 4 g, 4.5 g, 5 g,5.5 g, 6 g, 6.5g, 7 g, 7.5g, 8 g, 8.5 g, 9 g, 9.5 g, or 10 g of the oneor more second therapeutic agents herein.

In some embodiments, a composition herein comprises, consistsessentially of, or consists of 0.0001-10 g, 0.0005-9 g, 0.001-8 g,0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g of the one ormore second therapeutic agents herein.

Pharmaceutical Formulations

Pharmacologically acceptable aqueous vehicles for the compositions ofthe present invention can include, for example, any liquid solution thatis capable of dissolving a detergent and is not toxic to the particularindividual receiving the formulation. Examples of pharmaceuticallyacceptable aqueous vehicles include, without limitation, saline, waterand acetic acid. Typically, pharmaceutically acceptable aqueous vehiclesare sterile.

Pharmacologically active detergent compositions useful in embodiments ofthe present invention are formulated for the non-surgical reduction oflocalized fat deposits. As used herein, “non-surgical” refers to medicalprocedures that do not require an incision. Injections are examples ofnon-surgical procedures. Liposuction is a surgical procedure.

In one embodiment of the present invention, the pharmacologically activedetergent composition is administered by injection, for example, bybolus injection. In order to be effective, the detergent compositionmust have direct contact with the fat tissue regardless of how it isinfused. The detergent formulations can be injected subcutaneously orinfused directly into the fat. Formulations for injection can bepresented in unit dosage form, for example, in ampoules or in multi-dosecontainers, with an added preservative. The compositions can take suchforms as suspensions, solutions, or emulsions in oily or aqueousvehicles, and can contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

A “pharmaceutically acceptable excipient” may be used herein, and refersto a compound that is useful in preparing a pharmaceutical compositionthat is generally safe, non-toxic and neither biologically nor otherwiseundesirable, and includes excipients that are acceptable for veterinaryuse or human pharmaceutical use. A pharmaceutically acceptable excipientas used in the specification and claims includes both one and more thanone such excipient. Some examples of suitable excipients includelactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,calcium phosphate, alginates, tragacanth, gelatin, calcium silicate,microcrystalline cellulose, polyvinylpyrrolidone, phosphatidylcholine,cellulose, sterile water, syrup, and methyl cellulose. The formulationscan additionally include: lubricating agents such as talc, magnesiumstearate, and mineral oil; wetting agents; emulsifying and suspendingagents; and preserving agents such as methyl- andpropylhydroxy-benzoates and benzyl alcohol. The compositions of thepresent invention can be formulated so as to provide quick, sustained ordelayed release of the active ingredient after administration to thepatient by employing procedures known in the art.

Additional excipients suitable for formulation with the detergentcompositions of the present invention include penetration enhancers anddispersion agents. Non-limiting examples of dispersion agents whichallow the dispersion of drugs in tissue include hyaluronidase andcollagenase. Hyaluronidase functions to augment tissue permeability andspread or dispersion of other drugs. Collagenase has been used toisolate adipocytes from subcutaneous fat and does not have lytic effectson adipocytes themselves. Additionally hyaluronidase and collagenase canfacilitate healing by accelerating reduction of necrotic tissue aftertreatment with the detergent formulations of the present invention.

The pharmacologically active detergent compositions of the presentinvention are useful for treating localized fat accumulations, includingbut not limited to lower eyelid fat herniation, accumulations on thewaist, hips and other cosmetic areas, xanthelasma, lipomas andlipodistrophy, including “buffalo hump” lipodystrophy (3). In anotherembodiment, the detergent compositions of the present invention isuseful for treating fat deposits associated with cellulite.

Methods

The present invention also relates to methods for reducing asubcutaneous fat deposit in a mammal. Such methods comprise, consistessentially of, or consist of administering locally to the fat depositin the mammal one or more of the compositions or dose units herein. Forexample, in one embodiment less than 500 mL of a solution is deliveredlocally to the fat deposit to be reduced. The solution comprises,consists essentially of, or consists of pharmacologically activedetergent(s) (preferably bile salts such as sodium deoxycholate), suchas those disclosed herein. The solution preferably comprises less than5% w/v phosphatidylcholine or more preferably comprises nophosphatidylcholine.

In some embodiments of the present invention, the above methods areprovided for the non-surgical removal of one or more localized fatdeposits in a patient. For example, in one embodiment, the non-surgicalmethods herein do not include liposuction. In some embodiments, themethods herein also exclude non-invasive means for reducing fat, e.g.,ultrasonification. In other embodiments, non-invasive means can be usedin conjunction with the compositions herein.

The patient being treated is preferably a mammal. Such mammal can be ahuman or an animal such as a primate (e.g., a monkey, chimpanzee, etc.),a domesticated animal (e.g., a dog, cat, horse, etc.), farm animal(e.g., goat, sheep, pig, cattle, etc.), or laboratory animal (e.g.,mouse, rat, etc.). Preferably, a patient being treated is a human, ahorse, a dog, or a cat.

The compositions herein can be used to treat any adipose condition inthe patient including, for example, disorders such as lipomas,herniation, Dercum's disease, Madelung's neck, lipedema, piezogenicnodules, xanthelasma, lipodystrophy, and cellulite. In otherembodiments, the compositions herein can be used to treat adiposeconditions in areas such as fat deposits localized under eye, underchin, under arm, buttock, calf, back, thigh, ankle, or stomach of amammal.

The fat-solubilizing compositions herein are preferably administered viaa localized injection. However, other means of administering thecompositions herein are also contemplated. For example, the compositionsherein may be administered via a dermal patch or a subcutaneous depot.

Generally, the total volume, unit dose and number of treatmentsadministered will vary depending on the amount of fat in a target site,the location of the target site, type of fat composition, and desiredresults. In general, the greater the amount of fat being treated, thegreater the dose that is administered. It should be noted that while thecompositions and unit dosages herein may be administered into anindividual as part of a treatment regimen, they are not removed from theindividual as part of the treatment regimen.

Thus, the present invention contemplates methods for reducing amount ofsubcutaneous fat in a mammal by administering to the mammal an effectiveamount of a fat-solubilizing composition that comprises, consistsessentially of, or consists of one or more pharmacologically activedetergents. The above is preferably administered transdermally orsubcutaneously, via e.g., a subcutaneous injection using a syringe to atarget site. A target site can be for example 0.1 cm×0.1 cm, to about 5cm×5 cm. The compositions herein may be administered at the same,adjacent, or nearby target sites at various intervals, dosages, volumes,as disclosed herein.

The present invention provides compositions and methods for thenon-surgical reduction of localized fat deposits in a mammal. In oneembodiment, the methods herein involve administration offat-solubilizing concentrations one or more pharmacologically activedetergents in pharmaceutically acceptable injectable solutions. For thepurposes of the present invention, a non-surgical method of fatreduction does not include liposuction, lipoplasty or suction lipectomy.

Preferably, the methods herein exclude the non active removal of thepharmacologically active detergents (e.g., via suction).

Any of the above methods may be supplemented by further administering tothe patient a second therapeutic agent. The second therapeutic agent canbe administered separately or in combination with the compositionsherein. The second therapeutic agent can be administered locally orsystemically. In some embodiments, the second therapeutic agent isco-formulated with the detergent(s) and administered simultaneously withthe detergent(s) herein. In other one or more of the second therapeuticagents are administered prior to the administration of the detergentsherein.

The above may be administered once or multiple times into the targetsite. In some embodiments, the compositions herein are administered atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times to a target site. More than1 administration can occur in a single hour, day, week, month, or year.Preferably, multiple administrations into a single target site occurless than 10, 9, 8, 7, 6, 5, 4, 3, or two times a year, less than 10, 9,8, 7, 6, 5, 4, 3, or 2 times month, less than 10, 9, 8, 7, 6, 5, 4, 3,or two times a week, less than 10, 9, 8, 7, 6, 5, 4, 3, or two times aday or less than 10, 9, 8, 7, 6, 5, 4, 3, or two times an hour. In someembodiments a patient is given 1-100, 2-50, 3-30, 4-20, or 5-10injection at a target site. This number of injections can occur over aperiod of 1 year, 6 months, 5 months, 4 months, 3 months, 2 months, 1month, 3 weeks, 2, weeks, or 1 week or less.

The compositions can be administered at various levels below the dermis,including, for example, 0.1-4 inches, 0.5-3 inches, 1-2 inches below thedermis.

The compositions can be administered in various volumes but preferablyin a total volume of less than 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL,8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, 0.1 mL, 0.01 mL perinjection.

Kits

FIG. 7 is an illustration of a kit 101 for the reduction of subcutaneousfat accumulation in a mammal without the use of liposuction. The kitincludes one or more first containers 102. A first container 102comprises, consists essentially of, or consists of any of thecompositions herein. For example, a first container can comprise,consist essentially of, or consist of a pharmacologically activedetergent and less than 5% w/v phosphatidylcholine.

The above may be prepared in a solution or more preferably in aninjectable solution. First container(s) 102 comprising such a solutionmay have sufficient volume to hold one or more unit doses. For example,a first container 102 may be adapted to hold a less than 500 mL, 100 mLsolution, 20 mL solution 10 mL solution or 5 mL solution. In someembodiments, first container(s) 102 hold a volume of about 0.01 ml toabout 100 ml, about 0.1 ml to about 90 ml, about 0.5 ml to about 80 ml,about 1 ml to about 70 ml, about 2 ml to about 60 ml, about 3 ml toabout 50 ml, about 4 ml to about 40 ml, about 5 ml to about 30 ml, about6 ml to about 20 ml, and about 7 ml to about 10 ml. In more preferredembodiments, the first container(s) 102 is an ampule having a volumecapacity of about 10 to about 20 ml.

In some embodiments, the detergent and option phosphatidylcholine areformulated in a dermal patch or a depot for sustained release. Dosagesin a patch or depot can be the same as those discussed herein.

A first container 102 can optionally include one or more secondtherapeutic agents. Preferably, a first container 102 includes ananalgesic, antimicrobial agent, or anti-inflammatory agents. A firstcontainer 102 can also include a second detergent. Examples ofdetergents are described herein.

A first container 102 preferably has less than 5% w/v of phospholipids,such as phosphatidylcholine. In some embodiments, a first container 102contains no phospholipids or no phosphatidylcholine.

A first container 102 preferably has more than 0.01%, 0.1%, 1.0%, 2.0%,3.0%, 4.0%, or 5.0% w/w, w/v or v/v pharmacologically activedetergent(s). Preferably, the concentration of the pharmacologicallyactive detergent in the first container 102 is above its micellarconcentration. In some embodiments, the concentration of saidpharmacologically active detergent(s) in % w/v is greater than theconcentration of said phospholipids or phosphatidylcholine in % w/v.

The solution of container 102 is administered according to theinstructions for use 103. Instructions for use 103 can provide dosinginstructions which may depend upon, for example, target site, mammalbeing treated, desired results, location of target site, concentrationof solution, size of fat deposition. Preferably instructions for use 103are for the treatment of a mammal such as a human, a dog, a cat, or ahorse. Instructions for use 103 can also include information fortreatment of other domesticated animals and/or farm animals.

Instruction for use 103 may also include information on the use of thecompositions herein to treat specific target sites, such as, e.g., fatdeposits localized under eye, under chin, under arm, buttock, calf,back, thigh, ankle, or stomach of a mammal. In some embodiments,instruction for use 103 detail an explanation for use of thecompositions herein to treat a fat deposit that is eyelid fatherniation, lipomas, lipodystrophy, buffalo hump lipodystrophy or fatdeposits associated with cellulite.

Instruction for use 103 may include information regarding properdiluents and volumes for dilution, if any, of the first container 102and/or the second container 105. The instructions for use 103 may alsoprovide information regarding the proper administration of thecompositions herein, such as frequency and dosage of administration.

Kit 101 may further comprise a syringe or other suitable delivery device(e.g., patch, subcutaneous depot) 104 for delivering the compositions infirst container 102 to a subcutaneous fat accumulation region. In someembodiments, syringe or delivery device 104 may be preloaded with a unitdose of a solution of the present invention.

The kit 101 may further include a second container 105 comprising asecond active agent. Examples of a second therapeutic agent include, forexample, an antimicrobial agent, an anti-thrombotic agent, ananti-coagulation agent, a suds-depressant, an anti-inflammatory agent,an analgesic, an anesthetic, an anti-dispersion agent, a dispersionagent, a penetration enhancer, a steroid, a tranquilizer, a musclerelaxant, and an anti-diarrhea agent.

The following examples are provided to more precisely define and enablethe compositions and methods of the present invention. It is understoodthat there are numerous other embodiments and methods of using thepresent invention that will be apparent embodiments to those of ordinaryskill in the art after having read and understood this specification andexamples. The following examples are meant to illustrate one or moreembodiments of the invention and are not meant to limit the invention tothat which is described below.

Examples Example 1 Sodium Deoxycholate and PhosphatidylcholineFormulations

Phosphatidylcholine bile salt formulation (PBF) (5.0% highly purifiedsoy derived PC, 4.75% sodium deoxycholate, and 0.9% benzyl alcohol, insterile water, Table 2) was obtained from Hopewell Pharmacy, Hopewell,N.J. Sodium deoxycholate and Triton® X-100 detergent (Triton®, alkylarylpolyether alcohol) were obtained from Sigma-Aldrich Corp. (St. Louis,Mo.). Empigen® BB detergent (Empigen®, lauryldimethylbetaine,Calbiochem, Biosciences, Inc., La Jolla, Calif.). Stock reagents (5%dilutions) were prepared in PBS buffer.

The molecular structure of (a) phosphatidylcholine, (b) sodiumdeoxycholate and (c) benzyl alcohol are depicted in FIG. 1.

TABLE 2 Injectable PBF Phosphatidylcholine 5.00% (w/v) Sodiumdeoxycholate 4.75% Benzyl alcohol 0.90% Water 100 mL

Example 2 Effects of Sodium Deoxycholate and PhosphatidylcholineSolutions in Cultured Cells

To measure cell viability after detergent treatment, HaCaT humankeratinocyte cells were cultured in DMEM (Dulbecco's modified Eagle'smedium) supplemented with 10% fetal calf serum, penicillin, andstreptomycin. HaCaT cells were cultured in 6 well plates and incubatedwith 0%, 0.005%, 0.050% or 0.500% PBF (PC Formula) or sodiumdeoxycholate for 30 min at 37° C. prior to determination of cellviability using the MTS assay, which uses a tetrazolium compound thatproduces a color change when bioreduced by metabolically active cells(CellTiter 96® AQ_(ueous) Non-Radioactive Cell Proliferation Assay,Promega, Corp. Madison, Wis.). Cell viability was determined by anabsorbance spectrophotometer (at 490 nm) after a 4 hour incubation withthe assay at 37° C. To determine cell viability in fresh tissue, fatspecimens were incubated for 4 hours in 24 well plates with stockreagents and the MTS assay. Tissue specimens were then visualized forcolor change and the amount of MTS in their supernatants was measured byabsorbance (at 490 nm). All studies were performed in triplicate.Absorbance at 490 nm (OD 490) is proportional to the number of livingcells in the culture. There was comparable OD 490 in the control and0.005% dilutions of both compounds (FIG. 2 a), indicating little effectof these substances on cell viability at this concentration. Cellviability progressively decreased at 0.05% and 0.5% concentrations ofboth solutions.

Cell lysis in response to detergent treatment was determined in HaCaTcells incubated with the reagents at the indicated cell dilutions for 30min at 37° C. Lactate dehydrogenase release was measured by absorbance(at 490 nm) after a 1 hour incubation with the LDH assay as recommendedby the manufacturer (CytoTox 96® Non-Radioactive Cytotoxicity Assay,Promega). All studies were performed in triplicate. LDH release isdirectly proportional to absorbance at 490 nm (OD 490). There wasminimal LDH release from control cells and those incubated with 0.005%dilutions of both compounds (FIG. 2 b). There was progressively more LDHreleased at 0.05% and 0.5% of the PBF and deoxycholate.

Example 3 Effects of Sodium Deoxycholate and PhosphatidylcholineSolutions in Porcine Tissue

Porcine tissue was obtained immediately after sacrifice, shaved, andplaced on ice for a maximum of four hours before use. Fat specimens wereobtained by removing the epidermis and dermis of a punch biopsy with ascalpel and trimmed. Fat specimens were loaded with calcein dye byincubating 1 hour at 37° C. with Calcein-AM (Sigma). Stock reagents wereadded to the fat specimens and incubated for 30 min at 37° C. withgentle agitation. Calcein retention was determined by tissuefluorescence using purple (411 nm) light and visually observing theemitted green (500 nm) light using an emission filters.

Histology was performed by injecting stock reagent solutions (0.5 mL)into full thickness porcine skin at various levels (epidermis, dermis,and subcutaneous tissue) with 1.0 mL syringes and 30-gauge, 0.5 inchneedles. Needle depth was visualized along the margin of the porcinetissue with the intent of saturating the target tissue. One hour afterincubation with PBS at 37° C., multiple 5.0 mm biopsy specimens wereobtained from the injected sites, each condition performed intriplicate. Tissue was fixed in formaldehyde, paraffin-embedded, andstained with hematoxylin-eosin. Specimens were evaluated by aboard-certified dermatopathologist who was blinded to the treatmentprotocol.

Fresh porcine skin was used to determine if the effects of thesedetergent substances on cultured cells were similar in tissue. FIG. 3 ademonstrates the production of dark purple pigment (indicating viablecells) in fat tissue treated with the PBS buffer (negative control)using the MTS assay. The PBF and 5% solutions of deoxycholate andTriton® detergent (positive control) demonstrated a comparable loss ofpurple dye (indicating cell death) in the treated fat specimens. Thedifference in fat cell viability between the solutions was quantified bymeasuring the absorbance (at 490) of the supernatants collected from thetreated fat specimens (FIG. 3 b). All reagents had significant effectson the fat cell viability of fresh tissue.

Cell lysis was confirmed using a calcein dye release assay. Calceinbecomes fluorescent after hydrolysis and is retained in cells that haveintact cell membranes. Because it does not label dead cells and is lostunder conditions that cause cell lysis, loss of green fluorescence infat tissue samples loaded with the dye calcein indicates cell lysis(FIG. 4). Samples treated with the deoxycholate, PBF, and Triton®detergent (positive control) exhibited similar loss of fluorescence.

The histologic changes resulting from injection of PBF, deoxycholate,and Empigen®, are shown in FIG. 5. Phosphatidylcholine bile saltformulation (FIG. 5 b) and deoxycholate (FIG. 5 d) produced histologiceffects similar to those caused by Empigen® (FIG. 5 g) and Triton® (notshown), two well-characterized laboratory detergents. These changes wereapparent in both fat and muscle. Marked blurring and dissolution ofadipocyte cell membranes with disruption of its normal lobulararchitecture were seen, after injection of both the PBF (FIG. 5 b) anddeoxycholate (FIG. 5 d). FIG. 5 f demonstrates muscle fiber disarray andatrophy after PBF injection. Similar changes in muscle tissue werevisible in the specimens treated with deoxycholate and the Triton® andEmpigen® detergents. There were no changes in the epidermis, dermis, oradnexal structures after injection of the reagents with the exception ofEmpigen®, which caused loss of fibroblast nuclear staining andhyalinization of dermal collagen.

Example 4 Clinical Experience with Sodium Deoxycholate Compositions

Patients having lipomas, benign, isolated collections of adipose tissue,were injected with sodium deoxycholate (DC) solutions withoutphosphatidylcholine directly into the lipoma. The results of this studydemonstrate that the detergent effects of deoxycholate seen on fat inanimal tissues are reproducible clinically in humans. All injectedlipomas were reduced in size after at least one treatment with variedconcentrations of deoxycholate (Table 3). A lipoma from one patient,injected with 1% DC, was excised after treatment and pathological andhistological analysis performed. Within the excised lipoma, necrosis isvisible grossly (FIG. 6 a) with a well demarcated area of hemorrhage andnecrosis on the lateral edge extending into the middle of the lipoma fatwhich contrasts with the normal lipoma fat which is lighter in color.Histological analysis (FIG. 6 b) reveals a well defined area ofhemorrhage and necrotic fat as well as a significant inflammatoryreaction which contrasts to the adjacent normal round clear fat cells.

TABLE 3 Reduction in size of lipomas after DC treatment Lipoma Size (cm)Size (cm) Total Treatments Pre-treatment Post-treatment (% DC injected)1 2.00 × 1.00 1.25 × 0.50 2 (2.5%) 2 2.00 1.50 × 0.50 3 (5% and 2.5%) 32.00 × 2.50 2.00 × 1.00 3 (5% and 2.5%) 4 4.00 × 1.75 2.50 × 2.00 2 (1%)5 2.00 × 1.75 1.25 2 (1%) 6 2.80 0.50 1 (5%) 7 1.00 Imperceptible 1 (1%)

Unless otherwise indicated, all numbers expressing quantities ofingredients, properties such as molecular weight, reaction conditions,and so forth used in the specification and claims are to be understoodas being modified in all instances by the term “about.” Accordingly,unless indicated to the contrary, the numerical parameters set forth inthe following specification and attached claims are approximations thatmay vary depending upon the desired properties sought to be obtained bythe present invention. At the very least, and not as an attempt to limitthe application of the doctrine of equivalents to the scope of theclaims, each numerical parameter should at least be construed in lightof the number of reported significant digits and by applying ordinaryrounding techniques. Notwithstanding that the numerical ranges andparameters setting forth the broad scope of the invention areapproximations, the numerical values Size (cm) set forth in the specificexamples are reported as precisely as possible. Any numerical value,however, inherently contains certain errors necessarily resulting fromthe standard deviation found in their respective testing measurements.

The terms “a” and “an” and “the” and similar referents used in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. Recitation of ranges of values herein is merely intended toserve as a shorthand method of referring individually to each separatevalue falling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g. “such as”) provided herein isintended merely to better illuminate the invention and does not pose alimitation on the scope of the invention otherwise claimed. No languagein the specification should be construed as indicating any non-claimedelement essential to the practice of the invention.

Groupings of alternative elements or embodiments of the inventiondisclosed herein are not to be construed as limitations. Each groupmember may be referred to and claimed individually or in any combinationwith other members of the group or other elements found herein. It isanticipated that one or more members of a group may be included in, ordeleted from, a group for reasons of convenience and/or patentability.When any such inclusion or deletion occurs, the specification is hereindeemed to contain the group as modified thus fulfilling the writtendescription of all Markush groups used in the appended claims.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention. Ofcourse, variations on those preferred embodiments will become apparentto those of ordinary skill in the art upon reading the foregoingdescription. The inventor expects skilled artisans to employ suchvariations as appropriate, and the inventors intend for the invention tobe practiced otherwise than specifically described herein. Accordingly,this invention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

Furthermore, numerous references have been made to patents and printedpublications throughout this specification. Each of the above citedreferences and printed publications are herein individually incorporatedby reference in their entirety.

In closing, it is to be understood that the embodiments of the inventiondisclosed herein are illustrative of the principles of the presentinvention. Other modifications that may be employed are within the scopeof the invention. Thus, by way of example, but not of limitation,alternative configurations of the present invention may be utilized inaccordance with the teachings herein. Accordingly, the present inventionis not limited to that precisely as shown and described.

REFERENCES

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Structural and    kinetic studies on the solubilization of lecithin by sodium    deoxycholate. Biochemistry 1979, 18:3517-25.-   10. Lichtenberg D, Robson R J, Dennis E A. Solubilization of    phospholipids by detergents. Structural and kinetic aspects. Biochim    Biophys Acta 1983, 737:285-304.-   11. Teelmann K, Schlappi B, Schupbach M, Kistler A. Preclinical    safety evaluation of intravenously administered mixed micelles.    Arzneimittelforschung 1984, 34:1517-23.-   12. Durr M, Hager J, Lohr J P. Investigation on mixed micelle and    liposome preparations for parental use on soya phosphatidylcholine.    Eur J Pharm Biopharm 1994, 40:147-56.-   13. Alkan-Onyuksel H, Ramakrishnan S, Chai H B, Pezzuto J M. A mixed    micellar formulation suitable for the parenteral administration of    taxol. Pharm Res 1994, 11:206-12.-   14. Hammad M A, Muller B W. Increasing drug solubility by means of    bile salt-phosphatidylcholine-based mixed micelles. Eur J Pharm    Biopharm 1998, 46:361-7.-   15. Parnham M J, Wendel A. Phospholipids and liposomes—safety for    cosmetical and pharmaceutical use. Nattermann Phospholipid GMBH    Scientific Publication No. 2 1995.-   16. Lipostabil. Product insert: Aventis Pharma, 2003.-   17. Goldman L, Bennet J C, Cecil R L. Cecil Textbook of Medicine.    St. Louis, Mo.: W.B. Saunders Co., 2001.-   18. Womack M D, Kendall D A, MacDonald R C. Detergent effects on    enzyme activity and solubilization of lipid bilayer membranes.    Biochim Biophys Acta 1983, 733:210-5.-   19. Lichtenberg D. Characterization of the solubilization of lipid    bilayers by surfactants. Biochim Biophys Acta 1985, 821:470-8.-   20. Banerjee P, Joo J B, Buse J T, Dawson G. Differential    solubilization of lipids along with membrane proteins by different    classes of detergents. Chem Phys Lipids 1995, 77:65-78.-   21. Almgren M. Mixed micelles and other structures in the    solubilization of bilayer lipid membranes by surfactants. Biochim    Biophys Acta 2000, 1508:146-63.-   22. Schuck S, Honsho M, Ekroos K, Shevchenko A, Simons K. Resistance    of cell membranes to different detergents. Proc Natl Acad Sci 2003,    100:5795-800.-   23. Heerklotz H, Seelig J. Correlation of membrane/water partition    coefficients of detergents with the critical micelle concentration.    Biophys J 2000, 78:2435-40.-   24. Learn about lecithins. Oxford, Conn.: American Lecithin Company,    2003.-   25. Canty D, Zeisel S, Jolitz A. Lecithin and choline: research    update on health and nutrition. Fort Wayne, Ind.: Central Soya    Company, Inc., 1996.-   26. Feldman M, Scharschmidt B F, Sleisenger M H, Fordtran J S,    Zorab R. Sleisenger & Fordtran's Gastrointestinal and Liver Disease.    New York: Saunders, 2002.-   27. Lipostabil. Rhone-Polenc Rorer. Cologne, West Germany: Natterman    International GMBH, 1990.-   28. Jones M N. Surfactants in membrane solubilisation. Int J Pharm    1999, 177:137-59.-   29. Gustafson C, Tagesson C. Influence of organic solvent mixtures    on biological membranes. Br J Ind Med 1985, 42:591-5.-   30. Lester D S, Baumann D. Action of organic solvents on protein    kinase C. Eur J Pharmacol 1991, 206:301-8.-   31. Engelke M, Jessel R, Wiechmann A, Diehl H A. Effect of    inhalation anaesthetics on the phase behaviour, permeability and    order of phosphatidylcholine bilayers. Biophys Chem 1997, 67:127-38.-   32. Ebihara L, Hall J E, MacDonald R C, McIntosh T J, Simon S A.    Effect of benzyl alcohol on lipid bilayers. A comparisons of bilayer    systems. Biophys J 1979, 28:185-96.-   33. Gordon L M, Sauerheber R D, Esgate J A, Dipple I, Marchmont R J,    Houslay M D. The increase in bilayer fluidity of rat liver plasma    membranes achieved by the local anesthetic benzyl alcohol affects    the activity of intrinsic membrane enzymes. 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1-53. (canceled)
 54. A non-surgical method for improving the appearance of a human having a localized fat accumulation and desiring the removal thereof, said method comprises local administration to said fat accumulation a composition consisting essentially of a detergent and phosphatidylcholine, which detergent is deoxycholic acid, or a salt thereof, wherein said detergent reduces the localized fat accumulation thereby improving the appearance of said human, and wherein said detergent is contained in excess of phosphatidylcholine by mass.
 55. A non-surgical method for improving the appearance of a human having a localized fat accumulation and desiring the removal thereof, said method comprises local administration to said fat accumulation a composition comprising a detergent and phosphatidylcholine, which detergent is deoxycholic acid, or a salt thereof, wherein said detergent reduces the localized fat accumulation thereby improving the appearance of said human, and wherein said detergent is contained in excess of phosphatidylcholine by mass.
 56. The method of claim 54 or 55, wherein said fat accumulation is localized in an area selected from the group consisting of: a thigh, the abdomen, the buttock, calf, an ankle, the upper back, the chin, an arm, the waist, and the hip.
 57. A method for treating a patient having lipomas comprising local administration to said lipomas of said patient a composition consisting essentially of a detergent and phosphatidylcholine, which detergent is deoxycholic acid, or a salt thereof, wherein said detergent reduces the size of said lipomas thereby treating said patient, and wherein said detergent is contained in excess of phosphatidylcholine by mass.
 58. A method for treating a patient having lipomas comprising local administration to said lipomas of said patient a composition comprising a detergent and phosphatidylcholine, which detergent is deoxycholic acid, or a salt thereof, wherein said detergent reduces the size of said lipomas thereby treating said patient, and wherein said detergent is contained in excess of phosphatidylcholine by mass.
 59. The method of any one of claims 54, 57, and 58, wherein the concentration of the detergent is up to 5% (w/v) deoxycholic acid or the salt thereof.
 60. The method of any one of claims 54, 57, and 58, wherein the concentration of the detergent is about 0.1%, 0.5%, or 1% (w/v) deoxycholic acid or the salt thereof.
 61. The method of any one of claims 54, 57, and 58, wherein said salt is sodium salt.
 62. The method of any one of claims 54, 57, and 58, wherein said local administration comprises subcutaneous injection.
 63. The method of any one of claims 54, 57, and 58, wherein said composition further comprises a penetration enhancer.
 64. The method of any one of claims 54, 57, and 58, wherein said composition further comprises hyaluronidase and/or collagenase.
 65. The method of any one of claims 54, 57, and 58, wherein said composition further comprises an active ingredient which is not phosphatidylcholine.
 66. The method of claim 65, wherein said active ingredient is an anti-inflammatory agent and/or an analgesic.
 67. The method of claim 65, wherein said active agent is an analgesic.
 68. The method of claim 67, wherein said analgesic is lidocaine.
 69. The method of any one of claims 54, 57, and 58, wherein the ratio of the detergent to phosphatidylcholine is 1:0.5.
 70. The method of any one of claims 54, 57, and 58, wherein the ratio of the detergent to phosphatidylcholine is 1:0.05.
 71. The method of any one of claims 54, 57, and 58, wherein the ratio of the detergent to phosphatidylcholine is 1:0.005.
 72. A method for non-surgical removal of a localized fat deposit in a patient having such deposit and desiring to remove such deposit, which method comprises contacting the fat deposit with a composition comprising an effective amount of deoxycholic acid or a salt thereof, phosphatidylcholine and a pharmaceutically acceptable excipient, and wherein said deoxycholic acid or the salt thereof is contained in excess of phosphatidylcholine by mass.
 73. A method for non-surgical removal of a localized fat deposit in a patient having such deposit and desiring to remove such deposit, which method comprises contacting the fat deposit with a composition consisting essentially of an effective amount of deoxycholic acid or a salt thereof, phosphatidylcholine and a pharmaceutically acceptable excipient, and wherein said deoxycholic acid or the salt thereof is contained in excess of phosphatidylcholine by mass.
 74. A cosmetic method for non-surgical removal of a localized fat deposit in a patient having such deposit and desiring to remove such deposit, which method comprises contacting the fat deposit with a composition comprising an effective amount of deoxycholic acid or a salt thereof, phosphatidylcholine and a pharmaceutically acceptable excipient, and wherein said deoxycholic acid or the salt thereof is contained in excess of phosphatidylcholine by mass.
 75. A cosmetic method for non-surgical removal of a localized fat deposit in a patient having such deposit and desiring to remove such deposit, which method comprises contacting the fat deposit with a composition consisting essentially of an effective amount of deoxycholic acid or a salt thereof, phosphatidylcholine and a pharmaceutically acceptable excipient, and wherein said deoxycholic acid or the salt thereof is contained in excess of phosphatidylcholine by mass.
 76. A syringe for subcutaneous injection wherein said syringe comprises a sterile aqueous composition comprising a fat removing effective amount of up to 5% (w/v) of a detergent which is deoxycholic acid or a salt thereof, and phosphatidylcholine, wherein the detergent is contained in excess of phosphatidylcholine by mass.
 77. The syringe of claim 76, further containing at least another active ingredient.
 78. The syringe of claim 76, wherein the fat removing effective amount is from 0.05% to 5% (w/v).
 79. The syringe of claim 76, wherein the detergent is the salt of deoxycholic acid.
 80. The syringe of claim 76, wherein the salt is sodium deoxycholate.
 81. A composition comprising an adipose cell and aqueous solution of a fat removing bile acid or a salt thereof at a concentration of no more than 5% (w/v) and phosphatidylcholine, wherein the bile acid or the salt thereof is contained in excess of phosphatidylcholine by mass.
 82. The composition of claim 81, wherein the concentration of the bile acid or the salt is from 0.05% to 5% (w/v).
 83. The composition of claim 82, wherein the bile acid is deoxycholic acid.
 84. The composition of claim 82, wherein the bile acid salt is sodium deoxycholate. 